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Sample records for arthrobacter globiformis atcc

  1. Cloning and sequencing of the trpE gene from Arthrobacter globiformis ATCC 8010 and several related subsurface Arthrobacter isolates

    Energy Technology Data Exchange (ETDEWEB)

    Chernova, T.; Viswanathan, V.K.; Austria, N.; Nichols, B.P.

    1998-09-01

    Tryptophan dependent mutants of Arthrobacter globiformis ATCC 8010 were isolated and trp genes were cloned by complementation and marker rescue of the auxotrophic strains. Rescue studies and preliminary sequence analysis reveal that at least the genes trpE, trpC, and trpB are clustered together in this organism. In addition, sequence analysis of the entire trpE gene, which encodes component I of anthranilate synthase, is described. Segments of the trpE gene from 17 subsurface isolates of Arthrobacter sp. were amplified by PCR and sequenced. The partial trpE sequences from the various strains were aligned and subjected to phylogenetic analysis. The data suggest that in addition to single base changes, recombination and genetic exchange play a major role in the evolution of the Arthrobacter genome.

  2. Bioavailability of Ag(I) with Arthrobacter oxidas and Arthrobacter globiformis

    CERN Document Server

    Gelagutashvili, E; Ginturi, E; Bagdavadze, N; Kuchava, N; Tsakadze, K; Janjalia, M

    2012-01-01

    The biosorption of Ag(I)_ Arthrobacter species (Arthrobacter globiformis 151B and Arthrobacter oxidas 61B) was studied at simultaneous application of dialysis and atomic absorption analysis. The biosorption constants and nature of interaction for Ag(I) -Arthrobacter oxidas and Ag(I) -Arthrobacter globiformis were determined. The biosorption constants for Ag(I)- -Arthrobacter globiformis and for Ag(I) -Arthrobacter oxidas equal to 65.0 x10-4 and 35.0 x10-4 respectively.

  3. Electrochemical sensor based on Arthrobacter globiformis for cholinesterase activity determination.

    Science.gov (United States)

    Stoytcheva, Margarita; Zlatev, Roumen; Valdez, Benjamin; Magnin, Jean-Pierre; Velkova, Zdravka

    2006-07-15

    The sensors applied recently for determination of cholinesterase activity are mostly enzymatic amperometric sensors, in spite of their disadvantages: short life-time at ambient temperature, instability of the response, interferences, as well as passivation of the electrode surface. In the present paper a new approach for determination of cholinesterase activity was proposed, overcoming the main drawbacks of the analysis performed with amperometric enzymatic sensors. Instead of the immobilization of enzymes on a conducting electrode surface, whole cells of Arthrobacter globiformis, containing choline oxidase were fixed on a Clark type oxygen probe. Current proportional to bacteria respiration is registered as a sensor response. The application of whole cells of bacteria as a sensing element permits to achieve high stability of the response and long life-time of the sensor at ambient temperature, due to the conservation of the enzyme in its natural micro-environment inside the immobilized cells. The proposed sensor keeps its functionality more than 7 weeks stored in deionized water at ambient temperature. For the first 2 weeks the amplitude of the response decreases with only 10% and at the end of the studied 7 weeks period the response was 50% of the initial. The other advantages of the proposed sensor are: the dissolved oxygen is used as a mediator which concentration can be reliably and interferences free measured by the aim of a Clark type oxygen probe applied as a transducer; reproducible bacterial membranes can be elaborated by filtration of resuspended bacterial culture after preliminary determination of its activity; application of membranes containing lyophilized bacteria capable to be conserved infinitely long time and activated just before their application; negligible cost compared with the sensors based on immobilized enzymes. The steady-state response of the proposed bacterial sensor to choline obtained in 200 s is linear in the investigated

  4. Increased iron-stress resilience of maize through inoculation of siderophore-producing Arthrobacter globiformis from mine.

    Science.gov (United States)

    Sharma, Meenakshi; Mishra, Vandana; Rau, Nupur; Sharma, Radhey Shyam

    2016-07-01

    Iron deficiency is common among graminaceous crops. Ecologically successful wild grasses from iron-limiting habitats are likely to harbour bacteria which secrete efficient high-affinity iron-chelating molecules (siderophores) to solubilize and mobilize iron. Such siderophore-producing rhizobacteria may increase the iron-stress resilience of graminaceous crops. Considering this, 51 rhizobacterial isolates of Dichanthium annulatum from iron-limiting abandoned mine (∼84% biologically unavailable iron) were purified and tested for siderophore production; and efficacy of Arthrobacter globiformis inoculation to increase iron-stress resilience of maize and wheat was also evaluated. 16S rRNA sequence analyses demonstrated that siderophore-producing bacteria were taxonomically diverse (seven genera, nineteen species). Among these, Gram-positive Bacillus (eleven species) was prevalent (76.92%). A. globiformis, a commonly found rhizobacterium of graminaceous crops was investigated in detail. Its siderophore has high iron-chelation capacity (ICC: 13.0 ± 2.4 μM) and effectively dissolutes diverse iron-complexes (FeCl3 : 256.13 ± 26.56 μM/ml; Fe2 O3 red: 84.3 ± 4.74 μM/ml; mine spoil: 123.84 ± 4.38 μM/ml). Siderophore production (ICC) of A. globiformis BGDa404 also varied with supplementation of different iron complexes. In plant bioassay with iron-deficiency sensitive species maize, A. globiformis inoculation triggered stress-associated traits (peroxidase and proline) in roots, enhanced plant biomass, uptake of iron and phosphate, and protein and chlorophyll contents. However, in iron deficiency tolerant species wheat, growth improvement was marginal. The present study illustrates: (i) rhizosphere of D. annulatum colonizing abandoned mine as a "hotspot" of siderophore-producing bacteria; and (ii) potential of A. globiformis BGDa404 inoculation to increase iron-stress resilience in maize. A. globiformis BGDa404 has the potential to develop as

  5. Toxicity screening of soils from different mine areas—A contribution to track the sensitivity and variability of Arthrobacter globiformis assay

    Energy Technology Data Exchange (ETDEWEB)

    Marques, Catarina R., E-mail: crmarques@ua.pt [Departamento de Biologia and CESAM (Centro de Estudos do Ambiente e do Mar), Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro (Portugal); Caetano, Ana L. [Departamento de Biologia and CESAM (Centro de Estudos do Ambiente e do Mar), Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro (Portugal); Haller, Andreas [ECT Oekotoxikologie GmbH, Böttgerstraße 2–14, D-65439 Flörsheim a. M. (Germany); Gonçalves, Fernando [Departamento de Biologia and CESAM (Centro de Estudos do Ambiente e do Mar), Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro (Portugal); Pereira, Ruth [Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre, s/n, 4169-007 Porto (Portugal); Interdisciplinary Centre of Marine and Environmental Research (CIIMAR/CIMAR), Universidade do Porto, Rua dos Bragas 289, P 4050-123 Porto (Portugal); Römbke, Jörg [ECT Oekotoxikologie GmbH, Böttgerstraße 2–14, D-65439 Flörsheim a. M. (Germany)

    2014-06-01

    Highlights: • The assay gave rapid and feasible discrimination of toxic soils to A. globiformis. • Sensitive and low variability response to soils from different regions. • Soil properties may interfere with metal toxicity and fluorescence measurements. • Proposal of a toxicity threshold for the contact assay regarding soils. • A. globiformis assay should be included in the Tier I of risk assessment frameworks. - Abstract: This study used the Arthrobacter globiformis solid-contact test for assessing the quality of soils collected in areas subjected to past and present mine activities in Europe (uranium mine, Portugal) and North Africa (phosphogypsum pile, Tunisia; iron mine, Morocco). As to discriminate the influence of soils natural variability from the effect of contaminants, toxicity thresholds were derived for this test, based on the dataset of each study area. Furthermore, the test sensitivity and variability was also evaluated. As a result, soils that inhibited A. globiformis dehydrogenase activity above 45% or 50% relatively to the control, were considered to be toxic. Despite the soil metal content determined, the properties of soils seemed to influence dehydrogenase activity. Overall, the contact test provided a coherent outcome comparing to other more time-consuming and effort-demanding ecotoxicological assays. Our results strengthened the feasibility and ecological relevance of this assay, which variability was quite reduced hence suggesting its potential integration within the test battery of tier 1 of soil risk assessment schemes.

  6. Isolation of Arthrobacter Bacteriophage from Soil †

    OpenAIRE

    Germida, James J.; Casida, L. E.

    1981-01-01

    Soil was percolated with water and various nutrient solutions, and then the percolates were analyzed for bacteriophages which produced plaques on various Arthrobacter strains. The water percolates did not contain detectable phage. In contrast, phages for A. globiformis strains ATCC 8010 and 4336, and for several recent Arthrobacter species soil isolates, were easily detected in nutrient broth, soil extract, and cation-complete medium percolates. These percolates did not contain phage that pro...

  7. Degradation of the Phosphonate Herbicide Glyphosate by Arthrobacter atrocyaneus ATCC 13752

    OpenAIRE

    Pipke, Rüdiger; Amrhein, Nikolaus

    1988-01-01

    Of nine authentic Arthrobacter strains tested, only A. atrocyaneus ATCC 13752 was capable of using the herbicide glyphosate [N-(phosphonomethyl)glycine] as its sole source of phosphorus. Contrary to the previously isolated Arthrobacter sp. strain GLP-1, which degrades glyphosate via sarcosine, A. atrocyaneus metabolized glyphosate to aminomethylphosphonic acid. The carbon of aminomethylphosphonic acid was entirely converted to CO2. This is the first report on glyphosate degradation by a bacte...

  8. 球形节杆菌C224分批发酵2-酮基-D-葡萄糖酸的条件优化及其动力学模型的构建%Optimization of 2-keto-D-gluconic acid batch fermentation by Arthrobacter globiformis C224 and its kinetics

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    孙文敬; 高培玲; 魏转; 崔凤杰; 周延政; 滕文华; 刘敬泽

    2013-01-01

    In the present study,the operation parameters including aeration rate and initial glucose concentration during 50-L batch fermentation were optimized to increase the 2-keto-D-gluconic acid (2KGA) productivity from rice starch hydrolysate by Arthrobacter globiformis C224.The kinetic models for describing the cell growth,2KGA production and glucose consumption were proposed by using Logistic/Luedeking-Piret equations and mass balance calculation.Results showed that the 2KGA yield decreased slightly and its productivity lowered significantly when the aeration rate was below 1.5 v · v-1 · m-1 The initial glucose concentrations varying from 120.0 g/L to 180.0 g/L were optimal.Under the optimized conditions,the productivities and yields reached to 6.36 ~6.54 g/(L · h) and 0.96 ~0.97 g/g,respectively.Those proposed models could fit the predictive values well with the obtained experimental data.The determination coefficients (R2) of all models were over 0.95,which also indicated that theses models could elucidate the 2KGA metabolic characteristics of Arthrobacter globiformis C224.%通过优化发酵条件,提高球形节杆菌C224菌株利用大米淀粉水解糖分批发酵2-酮基-D-葡萄糖酸的生产强度,并在此基础上构建其发酵动力学模型.在50 L的发酵罐上考察了通气量与葡萄耱浓度对2-酮基-D-葡萄糖酸发酵的影响,并基于Logistic方程、Luedeking-Piret方程和物料平衡计算分别构建了菌体生长、产物形成和底物消耗的动力学模型.研究结果表明,通气量低于1.5 v·v-1·m-1时,发酵生产强度明显减小,糖酸转化率有所降低;发酵培养基中的葡萄糖浓度以120.0~180.0 g/L为宜,过高或过低对生产强度和糖酸转化率均有不利影响.在适宜的发酵条件下,球形节杆菌C224菌株分批发酵生产2-酮基-D-葡萄糖酸的生产强度达6.36~6.54 g/(L h),糖酸转化率为0.96 ~ 0.97 g/g.所构建动力学模型的计算值与实验值拟合效果

  9. MERCURY ADSORPTION BY ARTHOBACTER GLOBIFORMIS AND SPIRULINA PLATENSIS

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    T. L. Kalabegishvili

    2011-06-01

    Full Text Available The increasing contamination of soil, sediment, and water with heavy metals by natural and industrial processes is a worldwide problem. Many bacteria and microalgae have demonstrated ability to absorb toxic elements. To study mercury biosorption by bacteria Arthrobacter globiformis and microalga Spirulina platensis neutron activation analysis (NAA was applied. The process of mercury biosorption by these media was described by Freundlich and Langmuir-Freundlich Model. Both microorganisms showed a great potential to be used as biosorbing agents for mercury removal from the environment.

  10. Biosorption of Cr(VI)_ and Cr(III)_Arthrobacter species

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    Gelagutashvili, E.; Pataraia, E. Ginturi D.; Gurielidze, M.

    2011-01-01

    The biosorption of Cr(VI)_ and Cr(III)_ Arthrobacter species (Arthrobacter globiformis and Arthrobacter oxidas) was studied simultaneous application dialysis and atomic absorption analysis. Also biosorption of Cr(VI) in the presence of Zn(II) during growth of Arthrobacter species and Cr(III) in the presence of Mn(II) were discussed. Comparative Cr(VI)_ and Cr(III)_ Arthrobacter species shown, that Cr(III) was more effectively adsorbed by both bacterium than Cr(VI). The adsorption capacity is ...

  11. Ability of Cyanobacteria and Arthrobacter Species to Remove Gold Ions from Solution

    CERN Document Server

    Gelagutashvili, E; Rcheulichvili, A; Tsakadze, K; Bagdavadze, N; Kuchava, N; Djandjalia, M

    2012-01-01

    The biosorption of Au(III) - Spirulina platensis and Au(III) - Arthrobacter species (Arthrobacter globiformis and Arthrobacter oxidas) were studied at simultaneous application of dialysis and atomic absorption analysis. Also biosorption of Au(III) - Spirulina platensis at various pH were discussed. Biosorption constants for Au-cyanobacteris Spirulina platensis at different pH, and for Arthrobacter oxidas and Arthrobacter globiformis at pH=7.1 are : 1. K=3.91 x 10-4 (Au- Arthrobacter oxidas 61B, pH=7.1) 2. K=14.17 x 10-4 . (Au- Arthrobacter globiformis 151B, pH=7.1). 3. K=2.07x10-4 (Au- Spirulina platensis, pH=7.1) 4. K= 4.87x10-4 (Au- Spirulina platensis, pH=6.2) 5. K=8.7x10-4 (Au- Spirulina platensis, pH=8.4)

  12. Biosorption of Cr(VI)_ and Cr(III)_Arthrobacter species

    CERN Document Server

    Gelagutashvili, E; Gurielidze, M

    2011-01-01

    The biosorption of Cr(VI)_ and Cr(III)_ Arthrobacter species (Arthrobacter globiformis and Arthrobacter oxidas) was studied simultaneous application dialysis and atomic absorption analysis. Also biosorption of Cr(VI) in the presence of Zn(II) during growth of Arthrobacter species and Cr(III) in the presence of Mn(II) were discussed. Comparative Cr(VI)_ and Cr(III)_ Arthrobacter species shown, that Cr(III) was more effectively adsorbed by both bacterium than Cr(VI). The adsorption capacity is the same for both the Chromium-Arthrobacter systems. The biosorption constants for Cr(III) is higher than for Cr(VI) 5.7-5.9- fold for both species. Comparative Freundlich biosorption characteristics Cr(VI) Arthrobacter species of living and dry cells shown, that capacity(n) is in both cases the same(1.25,1.35). Dry cells have larger biosorption constant for both species, than living cells. Biosorption characteristics (K) and (n) for A. oxidas are without Mn(II) and in the presence of Mn(II) 2.6 x 10-4 (K), 1.37 (n) and 2...

  13. Uptake of Glyphosate by an Arthrobacter sp

    OpenAIRE

    Pipke, Rüdiger; Schulz, Arno; Amrhein, Nikolaus

    1987-01-01

    The uptake of glyphosate (N-[phosphonomethyl]glycine) by an Arthrobacter sp. which can utilize this herbicide as its sole source of phosphorus was investigated. Orthophosphate suppressed the expression of the uptake system for glyphosate and also competed with glyphosate for uptake. The Km for glyphosate uptake was 125 μM, and the Ki for orthophosphate was 24 μM. Organophosphonates as well as organophosphates inhibited glyphosate uptake, but only organophosphates and orthophosphate suppressed...

  14. Isolation of Arthrobacter species from the phyllosphere and demonstration of their epiphytic fitness

    OpenAIRE

    Scheublin, T.R.; Leveau, J.H.J.

    2013-01-01

    Bacteria of the genus Arthrobacter are common inhabitants of the soil environment, but can also be recovered from leaf surfaces (the phyllosphere). Using enrichment cultures on 4-chlorophenol, we succeeded in specifically isolating Arthrobacter bacteria from ground cover vegetation in an apple orchard. Based on 16S rRNA gene sequencing, the isolates were found to belong to at least three different species of Arthrobacter. Compared to the model bacterial epiphyte Pantoea agglomerans, the Arthr...

  15. Draft Genome Sequence of Arthrobacter chlorophenolicus Strain Mor30.16, Isolated from the Bean Rhizosphere

    OpenAIRE

    Miranda-Ríos, José Antonio; Ramírez-Trujillo, José Augusto; Nova-Franco, Bárbara; Lozano-Aguirre Beltrán, Luis Fernando; Iturriaga, Gabriel; Suárez-Rodríguez, Ramón

    2015-01-01

    Bacteria of the genus Arthrobacter are commonly found in the soil and plant rhizosphere. In this study we report the draft genome of Arthrobacter chlorophenolicus strain Mor30.16 that was isolated from rhizosphere of beans grown in Cuernavaca Morelos, Mexico. This strain promotes growth and ameliorates drought stress in bean plants.

  16. Wide Distribution of Closely Related, Antibiotic-Producing Arthrobacter Strains throughout the Arctic Ocean

    DEFF Research Database (Denmark)

    Wietz, Matthias; Månsson, Maria; Bowman, Jeff S.;

    2012-01-01

    We isolated 16 antibiotic-producing bacterial strains throughout the central Arctic Ocean, including seven Arthrobacter spp. with almost identical 16S rRNA gene sequences. These strains were numerically rare, as revealed using 454 pyrosequencing libraries. Arthrobacter spp. produced arthrobacilin...

  17. Adhesion of an Amylolytic Arthrobacter sp. to Starch-Containing Plastic Films

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    Imam, Syed H.; Gould, J. Michael

    1990-01-01

    Cells of the amylolytic bacterium KB-1 (thought to be an Arthrobacter sp.) adhered (∼70%) to the surface of plastic films composed of starch-poly (methylacrylate) graft copolymer (starch-PMA), but did not adhere (

  18. Plant compounds that induce polychlorinated biphenyl biodegradation by Arthrobacter sp. strain B1B.

    OpenAIRE

    Gilbert, E S; Crowley, D. E.

    1997-01-01

    Plant compounds that induced Arthrobacter sp. strain B1B to cometabolize polychlorinated biphenyls (PCBs) were identified by a screening assay based on the formation of a 4,4'-dichlorobiphenyl ring fission product. A chemical component of spearmint (Mentha spicata), l-carvone, induced Arthrobacter sp. strain B1B to cometabolize Aroclor 1242, resulting in significant degradation of 26 peaks in the mixture, including selected tetra- and pentachlorobiphenyls. Evidence for PCB biodegradation incl...

  19. Genome sequence of Arthrobacter antarcticus strain W2, isolated from a slaughterhouse

    DEFF Research Database (Denmark)

    Herschend, Jakob; Raghupathi, Prem Krishnan; Røder, Henriette Lyng;

    2016-01-01

    We report the draft genome sequence ofArthrobacter antarcticusstrain W2, which was isolated from a wall of a small slaughterhouse in Denmark. The 4.43-Mb genome sequence was assembled into 170 contigs.......We report the draft genome sequence ofArthrobacter antarcticusstrain W2, which was isolated from a wall of a small slaughterhouse in Denmark. The 4.43-Mb genome sequence was assembled into 170 contigs....

  20. Erythema caused by a localised skin infection with Arthrobacter mysorens

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    Chakraborty Trinad

    2010-12-01

    Full Text Available Abstract Background Skin erythemas of unknown origin are a frequent reason for consulting the general practitioner or dermatologist. Case presentation Here we report a case of an erythema resembling the erythema migrans manifestation of Lyme disease, but with atypical symptoms like persistent pruritus. The patient had no history of a recent tick-bite but displayed a positive serology for an advanced stage of Lyme borreliosis, which stood in contrast to the clinical manifestation of erythema migrans as a symptom of early Lyme disease. Three skin swabs and soil samples, collected in the area where the patient possibly acquired the infection, were examined by bacterial and fungal culture methods. Microorganisms were identified by using 16 S rRNA gene sequencing and bioinformatics. The patient and soil isolates were compared by employing RAPD analysis. The serum samples of the patient were examined by immunoblotting. Arthrobacter mysorens, a soil bacterium, was isolated from the collected skin and soil samples. The identity of both isolates was determined by molecular fingerprinting methods. A. mysorens was proven to be causative for the erythema by direct isolation from the affected skin and a positive serology, thus explaining the atypical appearance of the erythema compared to erythema migrans caused by Borrelia infection. Conclusions Infections with A.mysorens might be underreported and microbiological diagnostic techniques should be applied in cases of patients with unclear erythemas, resembling erythema migrans, without a history of tick bites.

  1. Application of NAA Method to Study Chromium Uptake by Arthrobacter oxydans

    CERN Document Server

    Tsibakhashvili, N Ya; Kalabegishvili, T L; Kirkesali, E I; Frontasyeva, M V; Pomyakushina, E V; Pavlov, S S

    2002-01-01

    To study chromium uptake by Arthrobacter oxydans (Cr(VI)-reducer bacteria isolated from Columbia basalt rocks, USA) instrumental neutron activation analysis method was applied. It was established that chromate accumulation is dose-dependent and it is more intesive in the interval of concentrations of Cr(VI) (10-50 mg/l). At low concentrations of Cr(VI) (up to 50 mg/l) the most intensive formation of Cr(V) was also found (using ESR method). Besides, it was estimated that reduction from Cr(VI) to Cr(V) is faster process than the uptake of Cr(VI). According to ENAA measurements Cr(III), in constant to Cr(VI), is not accumulated in Arthrobacter oxydans cells up to concentration of 200 mg/l. Using epithermal neutron activation analysis the background levels of 17 major, minor and trace elements were determined in Arthrobacter oxydans.

  2. Potential Degradation of Swainsonine by Intracellular Enzymes of Arthrobacter sp. HW08

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    Haili Li

    2013-11-01

    Full Text Available Swainsonine (SW is a toxin produced by locoweeds and harmful to the livestock industry. Degrading SW by Arthrobacter sp. HW08 was demonstrated as a promising way to deal with SW poisoning. However, it is unknown which part of the subcellular enzymes in Arthrobacter sp. HW08 is responsible for biodegrading SW and whether the metabolites are atoxic. In this study, intracellular and extracellular enzymes of Arthrobacter sp. HW08 were isolated and their enzyme activity was evaluated. The metabolites were fed to mice, and physiological and histological properties of the treated mice were investigated. The results showed that only intracellular enzyme of Arthrobacter sp. HW08 (IEHW08 could degrade SW efficiently. Compared with mice in SW treatment group, mice in SW + IEHW08 treatment group (1 increased their body weights; (2 showed higher number of platelets and lower number of white blood cells; (3 decreased the levels of creatinine, urea nitrogen, alanine transaminase and aspartate aminotransferase in serum; (4 reduced the number of vacuolated cells in cerebellum, liver and kidney. All these data demonstrate that IEHW08 was potentially safe for mice, while keeping the capacity of degrading SW. This study indicates a possible application of IEHW08 as an additive in the livestock industry to protect animals from SW poisoning.

  3. Arthrobacter luteus restriction endonuclease cleavage map of X174 RF DNA

    NARCIS (Netherlands)

    Vereijken, J.M.; Mansfeld, A.D.M. van; Baas, P.D.; Jansz, H.S.

    1975-01-01

    Cleavage of X174 RF DNA with the restriction endonuclease from Arthrobacter luteus (Alu I) produces 23 fragments of approximately 24–1100 base pairs in length. The order of most of these fragments has been established by digestion of Haemophilus influenzae Rd (Hind II) and Haemophilus aegyptius (Hae

  4. [Heterotrophic Nitrification and Aerobic Denitrification of the Hypothermia Aerobic Denitrification Bacterium: Arthrobacter arilaitensis].

    Science.gov (United States)

    He, Teng-xia; Ni, Jiu-pai; Li, Zhen-lun; Sun, Quan; Ye Qing; Xu, Yi

    2016-03-15

    High concentrations of ammonium, nitrate and nitrite nitrogen were employed to clarify the abilities of heterotrophic nitrification and aerobic denitrification of Arthrobacter arilaitensis strain Y-10. Meanwhile, by means of inoculating the strain suspension into the mixed ammonium and nitrate, ammonium and nitrite nitrogen simulated wastewater, we studied the simultaneous nitrification and denitrification ability of Arthrobacter arilaitensis strain Y-10. In addition, cell optical density was assayed in each nitrogen removal process to analyze the relationship of cell growth and nitrogen removal efficiency. The results showed that the hypothermia denitrification strain Arthrobacter arilaitensis Y-10 exhibited high nitrogen removal efficiency during heterotrophic nitrification and aerobic denitrification. The ammonium, nitrate and nitrite removal rates were 65.0%, 100% and 61.2% respectively when strain Y-10 was cultivated for 4 d at 15°C with initial ammonium, nitrate and nitrite nitrogen concentrations of 208.43 mg · L⁻¹, 201.16 mg · L⁻¹ and 194.33 mg · L⁻¹ and initial pH of 7.2. Nitrite nitrogen could only be accumulated in the medium containing nitrate nitrogen during heterotrophic nitrification and aerobic denitrification process. Additionally, the ammonium nitrogen was mainly removed in the inorganic nitrogen mixed synthetic wastewater. In short, Arthrobacter arilaitensis Y-10 could conduct nitrification and denitrification effectively under aerobic condition and the ammonium nitrogen removal rate was more than 80.0% in the inorganic nitrogen mixed synthetic wastewater. PMID:27337904

  5. The arthrobacter arilaitensis Re117 genome sequence reveals its genetic adaptation to the surface of cheese.

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    Christophe Monnet

    Full Text Available Arthrobacter arilaitensis is one of the major bacterial species found at the surface of cheeses, especially in smear-ripened cheeses, where it contributes to the typical colour, flavour and texture properties of the final product. The A. arilaitensis Re117 genome is composed of a 3,859,257 bp chromosome and two plasmids of 50,407 and 8,528 bp. The chromosome shares large regions of synteny with the chromosomes of three environmental Arthrobacter strains for which genome sequences are available: A. aurescens TC1, A. chlorophenolicus A6 and Arthrobacter sp. FB24. In contrast however, 4.92% of the A. arilaitensis chromosome is composed of ISs elements, a portion that is at least 15 fold higher than for the other Arthrobacter strains. Comparative genomic analyses reveal an extensive loss of genes associated with catabolic activities, presumably as a result of adaptation to the properties of the cheese surface habitat. Like the environmental Arthrobacter strains, A. arilaitensis Re117 is well-equipped with enzymes required for the catabolism of major carbon substrates present at cheese surfaces such as fatty acids, amino acids and lactic acid. However, A. arilaitensis has several specificities which seem to be linked to its adaptation to its particular niche. These include the ability to catabolize D-galactonate, a high number of glycine betaine and related osmolyte transporters, two siderophore biosynthesis gene clusters and a high number of Fe(3+/siderophore transport systems. In model cheese experiments, addition of small amounts of iron strongly stimulated the growth of A. arilaitensis, indicating that cheese is a highly iron-restricted medium. We suggest that there is a strong selective pressure at the surface of cheese for strains with efficient iron acquisition and salt-tolerance systems together with abilities to catabolize substrates such as lactic acid, lipids and amino acids.

  6. Isolation, purification and characterization of xylanase produced by Arthrobacter sp. MTCC 5214 when grown in solid-state fermentation

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    Khandeparker, R.; Bhosle, N.B.

    Thermoalkalophilic Arthrobacter sp. produced extracellular xylanase, when wheat bran, rice husk, rice bran and bagassae were used as carbon source under solid-state fermentation (SSF). The xylanase enzyme was isolated by ammonium sulfate (80...

  7. An Arthrobacter spp. bacteremia leading to fetal death and maternal disseminated intravascular coagulation.

    Science.gov (United States)

    Shigeta, Naoya; Ozaki, Kimiaki; Hori, Kensuke; Ito, Kimihiko; Nakayama, Masahiro; Nakahira, Kumiko; Yanagihara, Itaru

    2013-02-01

    A 34-year-old parous woman developed high fever and threatened preterm labor after a 1-day trip, for which she was receiving prenatal care at a hospital. Three days after onset, at 24 4/7 weeks of gestation, she was transferred to our hospital in an emergency. Soon after the woman's arrival at our hospital, the infant was spontaneously stillborn via a transvaginal delivery. Laboratory tests revealed severe maternal disseminated intravascular coagulation with renal and liver insufficiency. Histopathologic examination of the placenta revealed vast fibrin deposition and remarkable neutrophilic infiltration in the intervillous space, suggesting a rare bacterial infection caused by Arthrobacter spp. The bacteria were predominantly detected in the placenta and maternal blood serum by common bacterial 16S rRNA sequencing after polymerase chain reaction amplification. We report the first case, to our knowledge, of bacteremia with Arthrobacter spp., which may lead to maternal disseminated intravascular coagulation and intrauterine fetal death.

  8. Manganese transport in Brevibacterium ammoniagenes ATCC 6872.

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    Schmid, J.; Auling, G

    1987-01-01

    Uptake of manganese by Brevibacterium ammoniagenes ATCC 6872 was energy dependent and obeyed saturation kinetics (Km = 0.65 microM; Vmax = 0.12 mumol/min per g [dry weight]). Uptake showed optima at 27 degrees C and pH 9.5. 54Mn2+ accumulated by the cells was released by treatment with toluene or by exchange for unlabeled manganese ions, via an energy-dependent process. Co2+, Fe2+, Cd2+, and Zn2+ inhibited manganese uptake. Inhibition by Cd2+ and Zn2+ was competitive (Ki = 0.15 microM Cd2+ an...

  9. Improved biosorption for Cr(VI) reduction and removal by Arthrobacter viscosus using zeolite

    OpenAIRE

    Silva, Bruna Andreia Nogueira Airosa; Figueiredo, Hugo; Quintelas, C.; Neves, Isabel C.; Tavares, M.T.

    2012-01-01

    The aim of the present work was to optimize the reduction and removal of chromium from aqueous solutions by a biosorption system consisting of a bacteria supported on a zeolite. The system proposed combines the biosorption properties of Arthrobacter viscosus, with the ion exchange capacity of NaY zeolite. Experiments were also performed without the zeolite for comparison purposes. Experimental parameters such as solution pH, biomass concentration and initial Cr(VI) concentration were investig...

  10. Isolation and Preliminary Characterization of Twenty Bacteriophages Infecting Either Brevibacterium or Arthrobacter Strains

    OpenAIRE

    Trautwetter, Annie; Blanco, Carlos

    1988-01-01

    Thirty-seven bacteriophages plaquing on Corynebacterium, Brevibacterium, or Arthrobacter strains were isolated from soil or vegetation samples. Restriction analysis of phage DNA indicated that 20 phages were unique; one of them produced entirely turbid plaques on Brevibacterium ketoglutamicum and was characterized as temperate. All these phages were assigned to group B of the classification of Bradley (Bacteriol. Rev. 31:230-314, 1967) and had relatively narrow host ranges.

  11. Improving the Survival of Arthrobacter sp., CW9 during Spray Drying Monitored by Scan Electric Microscope

    OpenAIRE

    Zhenqiang Xia; Ming Zhu; Yanqiu Zhang

    2014-01-01

    The culture of an aquaculture probiotic, i.e., Arthrobacter sp., CW9, was spray dried with different carriers/protectants, in which Scan Electric Microscope (SEM) was used to analyze the surface of micro-paticles produced by spray-drying. Matrix of protectants, inlet temperature and feed rate were optimized according to the survival rate after spray drying. Scanning electron micrographs showed that cracks formed on the particle surface were a key factor in enhancing bacteria survival during s...

  12. Cesium and strontium tolerant Arthrobacter sp. strain KMSZP6 isolated from a pristine uranium ore deposit.

    Science.gov (United States)

    Swer, Pynskhem Bok; Joshi, Santa Ram; Acharya, Celin

    2016-12-01

    Arthrobacter sp. KMSZP6 isolated from a pristine uranium ore deposit at Domiasiat located in North-East India exhibited noteworthy tolerance for cesium (Cs) and strontium (Sr). The strain displayed a high minimum inhibitory concentration (MIC) of 400 mM for CsCl and for SrCl2. Flow cytometric analysis employing membrane integrity indicators like propidium iodide (PI) and thiazole orange (TO) indicated a greater sensitivity of Arthrobacter cells to cesium than to strontium. On being challenged with 75 mM of Cs, the cells sequestered 9612 mg Cs g(-1) dry weight of cells in 12 h. On being challenged with 75 mM of Sr, the cells sequestered 9989 mg Sr g(-1) dry weight of cells in 18 h. Heat killed cells exhibited limited Cs and Sr binding as compared to live cells highlighting the importance of cell viability for optimal binding. The association of the metals with Arthrobacter sp. KMSZP6 was further substantiated by Field Emission-Scanning Electron Microscopy (FE-SEM) coupled with Energy dispersive X-ray (EDX) spectroscopy. This organism tolerated up to 1 kGy (60)Co-gamma rays without loss of survival. The present report highlights the superior tolerance and binding capacity of the KMSZP6 strain for cesium and strontium over other earlier reported strains and reveals its potential for bioremediation of nuclear waste. PMID:27620733

  13. New metabolic pathway for degradation of 2-nitrobenzoate by Arthrobacter sp. SPG

    Science.gov (United States)

    Arora, Pankaj K.; Sharma, Ashutosh

    2015-01-01

    Arthrobacter sp. SPG utilized 2-nitrobenzoate as its sole source of carbon and energy and degraded it with accumulation of stoichiometric amounts of nitrite ions. Salicylate and catechol were detected as metabolites of the 2-nitrobenzoate degradation using high performance liquid chromatography and gas chromatography–mass spectrometry. Enzyme activities for 2-nitrobenzoate-2-monooxygenase, salicylate hydroxylase, and catechol-1,2-dioxygenase were detected in the crude extracts of the 2-nitrobenzoate-induced cells of strain SPG. The 2-nitrobenzoate-monooxygenase activity resulted in formation of salicylate and nitrite from 2-nitrobenzoate, whereas salicylate hydroxylase catalyzed the conversion of salicylate to catechol. The ring-cleaving enzyme, catechol-1,2-dioxygenase cleaved catechol to cis,cis-muconic acid. Cells of strain SPG were able to degrade 2-nitrobenzoate in sterile as well as non-sterile soil microcosms. The results of microcosm studies showed that strain SPG degraded more than 90% of 2-nitrobenzoate within 10–12 days. This study clearly shows that Arthrobacter sp. SPG degraded 2-nitrobenzoate via a new pathway with formation of salicylate and catechol as metabolites. Arthrobacter sp. SPG may be used for bioremediation of 2-nitrobenzoate-contaminated sites due to its ability to degrade 2-nitrobenzoate in soil. PMID:26082768

  14. New metabolic pathway for degradation of 2-nitrobenzoate by Arthrobacter sp. SPG

    Directory of Open Access Journals (Sweden)

    Pankaj Kumar Arora

    2015-06-01

    Full Text Available Arthrobacter sp. SPG utilized 2-nitrobenzoate as its sole source of carbon and energy and degraded it with accumulation of stoichiometric amounts of nitrite ions. Salicylate and catechol were detected as metabolites of the 2-nitrobenzoate degradation using high performance liquid chromatography and gas chromatography-mass spectrometry. Enzyme activities for 2-nitrobenzoate-2-monooxygenase, salicylate hydroxylase, and catechol-1,2-dioxygenase were detected in the crude extracts of the 2-nitrobenzoate-induced cells of strain SPG. The 2-nitrobenzoate-monooxygenase activity resulted in formation of salicylate and nitrite from 2-nitrobenzoate whereas salicylate hydroxylase catalyzed the conversion of salicylate to catechol. The ring-cleaving enzyme, catechol-1,2-dioxygenase cleaved catechol to cis, cis-muconic acid. Cells of strain SPG were able to degrade 2-nitrobenzoate in sterile as well as non-sterile soil microcosms. The results of microcosm studies showed that strain SPG degraded more than 90% of 2-nitrobenzoate within 10-12 days. This study clearly shows that Arthrobacter sp. SPG degraded 2-nitrobenzoate via a new pathway with formation of salicylate and catechol as metabolites. Arthrobacter sp. SPG may be used for bioremediation of 2-nitrobenzoate-contaminated sites due to its ability to degrade 2-nitrobenzoate in soil.

  15. Biosorption of Lead(II) by Arthrobacter sp. 25: Process Optimization and Mechanism.

    Science.gov (United States)

    Jin, Yu; Wang, Xin; Zang, Tingting; Hu, Yang; Hu, Xiaojing; Ren, Guangming; Xu, Xiuhong; Qu, Juanjuan

    2016-08-28

    In the present work, Arthrobacter sp. 25, a lead-tolerant bacterium, was assayed to remove lead(II) from aqueous solution. The biosorption process was optimized by response surface methodology (RSM) based on the Box-Behnken design. The relationships between dependent and independent variables were quantitatively determined by second-order polynomial equation and 3D response surface plots. The biosorption mechanism was explored by characterization of the biosorbent before and after biosorption using atomic force microscopy (AFM), scanning electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The results showed that the maximum adsorption capacity of 9.6 mg/g was obtained at the initial lead ion concentration of 108.79 mg/l, pH value of 5.75, and biosorbent dosage of 9.9 g/l (fresh weight), which was close to the theoretically expected value of 9.88 mg/g. Arthrobacter sp. 25 is an ellipsoidalshaped bacterium covered with extracellular polymeric substances. The biosorption mechanism involved physical adsorption and microprecipitation as well as ion exchange, and functional groups such as phosphoryl, hydroxyl, amino, amide, carbonyl, and phosphate groups played vital roles in adsorption. The results indicate that Arthrobacter sp. 25 may be potentially used as a biosorbent for low-concentration lead(II) removal from wastewater. PMID:27197671

  16. Draft Genome Sequence of Type Strain Streptococcus gordonii ATCC 10558

    DEFF Research Database (Denmark)

    Rasmussen, Louise Hesselbjerg; Dargis, Rimtas; Christensen, Jens Jørgen Elmer;

    2016-01-01

    Streptococcus gordonii ATCC 10558T was isolated from a patient with infective endocarditis in 1946 and announced as a type strain in 1989. Here, we report the 2,154,510-bp draft genome sequence of S. gordonii ATCC 10558T. This sequence will contribute to knowledge about the pathogenesis of...

  17. Quantification of the Effects of Salt Stress and Physiological State on Thermotolerance of Bacillus cereus ATCC 10987 and ATCC 14579

    NARCIS (Netherlands)

    Besten, den H.M.W.; Mataragas, M.; Moezelaar, R.; Abee, T.; Zwietering, M.H.

    2006-01-01

    The food-borne pathogen Bacillus cereus can acquire enhanced thermal resistance through multiple mechanisms. Two Bacillus cereus strains, ATCC 10987 and ATCC 14579, were used to quantify the effects of salt stress and physiological state on thermotolerance. Cultures were exposed to increasing concen

  18. Regulation and function of transaldolase isoenzymes involved in sugar and one-carbon metabolism in the ribulose monophosphate cycle methylotroph Arthrobacter P1

    NARCIS (Netherlands)

    Levering, P.R.; Dijkhuizen, Lubbert

    1986-01-01

    In the facultative methylotroph Arthrobacter P1 the enzyme transaldolase plays an important role in both the pentose phosphate pathway and in the ribulose monophosphate cycle of formaldehyde fixation. Among gluconate-negative mutants of Arthrobacter P1 strains occurred which also were unable to grow

  19. 2,3-Dihydroxybiphenyl dioxygenase gene was first discovered in Arthrobacter sp. strain P J3

    Institute of Scientific and Technical Information of China (English)

    YANG MeiYing; MA PengDa; LI WenMing; LIU JinYing; LI Liang; ZHU XiaoJuan; WANG XingZhi

    2007-01-01

    Bacterium strain PJ3, isolated from wastewater and identified as Arthrobacter sp. bacterium based on its 16S rDNA gene, could use carbazole as the sole carbon, nitrogen and energy source. The genomic libraryof strain PJ3 was constructed and a positive clone JM109 (pUCW402) was screened out for the expression of dioxygenase by the ability to form yellow ring-fission product. A 2,3-dihydroxybiphenyl dioxygenase (23DHBD) gene of 933 bp was found in the 3360 bp exogenous fragment of pUCW402 by GenSCAN software and BLAST analysis. The phylogenetic analysis showed that 23DHBD from strain PJ3 formed a deep branch separate from a cluster containing most known 23DHBD in GenBank.Southern hybridization confirmed for the first time that the 23DHBD gene was from the genomic DNA of Arthrobacter sp. PJ3. In order to test the gene function, recombinant bacterium BL21 (pETW-8) was constructed to express 23DHBD. The expression level in BL21 (pETW-8) was highest compared with the recombinant bacteria JM109 (pUCW402) and strain PJ3. We observed that 23DHBD was not absolute specific. The enzyme activity was higher with 2,3-dihydroxybiphenyl as a substrate than with catechol.The substrate specificity assay suggested that 23DHBD was essential for cleavage of bi-cyclic aromatic compounds during the course of aromatic compound biodegradation in Arthrobacter sp. strain PJ3.

  20. Complete genome sequence and metabolic potential of the quinaldine-degrading bacterium Arthrobacter sp. Rue61a

    Directory of Open Access Journals (Sweden)

    Niewerth Heiko

    2012-10-01

    Full Text Available Abstract Background Bacteria of the genus Arthrobacter are ubiquitous in soil environments and can be considered as true survivalists. Arthrobacter sp. strain Rue61a is an isolate from sewage sludge able to utilize quinaldine (2-methylquinoline as sole carbon and energy source. The genome provides insight into the molecular basis of the versatility and robustness of this environmental Arthrobacter strain. Results The genome of Arthrobacter sp. Rue61a consists of a single circular chromosome of 4,736,495 bp with an average G + C content of 62.32%, the circular 231,551-bp plasmid pARUE232, and the linear 112,992-bp plasmid pARUE113 that was already published. Plasmid pARUE232 is proposed to contribute to the resistance of Arthrobacter sp. Rue61a to arsenate and Pb2+, whereas the linear plasmid confers the ability to convert quinaldine to anthranilate. Remarkably, degradation of anthranilate exclusively proceeds via a CoA-thioester pathway. Apart from quinaldine utilization, strain Rue61a has a limited set of aromatic degradation pathways, enabling the utilization of 4-hydroxy-substituted aromatic carboxylic acids, which are characteristic products of lignin depolymerization, via ortho cleavage of protocatechuate. However, 4-hydroxyphenylacetate degradation likely proceeds via meta cleavage of homoprotocatechuate. The genome of strain Rue61a contains numerous genes associated with osmoprotection, and a high number of genes coding for transporters. It encodes a broad spectrum of enzymes for the uptake and utilization of various sugars and organic nitrogen compounds. A. aurescens TC-1 is the closest sequenced relative of strain Rue61a. Conclusions The genome of Arthrobacter sp. Rue61a reflects the saprophytic lifestyle and nutritional versatility of the organism and a strong adaptive potential to environmental stress. The circular plasmid pARUE232 and the linear plasmid pARUE113 contribute to heavy metal resistance and to the ability to degrade

  1. Impact of phenolic substrate and growth temperature on the arthrobacter chlorophenolicus proteome

    Energy Technology Data Exchange (ETDEWEB)

    Unell, Maria; Abraham, Paul E.; Shah, Manesh; Zhang, Bing; Ruckert, Christian; VerBerkmoes, Nathan C.; Jansson, Janet K.

    2009-02-15

    We compared the Arthrobacter chlorophenolicus proteome during growth on 4-chlorophenol, 4-nitrophenol or phenol at 5 C and 28 C; both for the wild type and a mutant strain with mass spectrometry based proteomics. A label free workflow employing spectral counting identified 3749 proteins across all growth conditions, representing over 70% of the predicted genome and 739 of these proteins form the core proteome. Statistically significant differences were found in the proteomes of cells grown under different conditions including differentiation of hundreds of unknown proteins. The 4-chlorophenol-degradation pathway was confirmed, but not that for phenol.

  2. Accumulation of rare earth elements by siderophore-forming Arthrobacter luteolus isolated from rare earth environment of Chavara, India

    Indian Academy of Sciences (India)

    E S Challaraj Emmanuel; T Ananthi; B Anandkumar; S Maruthamuthu

    2012-03-01

    In this study, Arthrobacter luteolus, isolated from rare earth environment of Chavara (Quilon district, Kerala, India), were found to produce catechol-type siderophores. The bacterial strain accumulated rare earth elements such as samarium and scandium. The siderophores may play a role in the accumulation of rare earth elements. Catecholate siderophore and low-molecular-weight organic acids were found to be present in experiments with Arthrobacter luteolus. The influence of siderophore on the accumulation of rare earth elements by bacteria has been extensively discussed.

  3. Genetic, morphological and physiological relationships among coryneform bacteria

    NARCIS (Netherlands)

    Crombach, W.H.J.

    1974-01-01

    The DNA base composition of the soil arthrobacters tested (65.3 - 67.0% GC) suggests that this group is genetically homogeneous. Hybridization experiments, however, revealed clear differences between the Arthrobacter simplex and the Arthrobacter globiformis strains. The orange cheese coryneforms wer

  4. Heavy metal resistance in Arthrobacter ramosus strain G2 isolated from mercuric salt-contaminated soil

    International Nuclear Information System (INIS)

    Present study describes isolation of a multiple metal-resistant Arthrobacter ramosus strain from mercuric salt-contaminated soil. The isolate was found to resist and bioaccumulate several metals, such as cadmium, cobalt, zinc, chromium and mercury. Maximum tolerated concentrations for above metals were found to be 37, 525, 348, 1530 and 369 μM, respectively. The isolate could also reduce and detoxify redox-active metals like chromium and mercury, indicating that it has great potential in bioremediation of heavy metal-contaminated sites. Chromate reductase and mercuric reductase (MerA) activities in protein extract of the culture were found to be 2.3 and 0.17 units mg-1 protein, respectively. MerA enzyme was isolated from the culture by (NH4)2SO4 precipitation followed by dye affinity chromatography and its identity was confirmed by nano-LC-MS/MS. Its monomeric molecular weight, and optimum pH and temperature were 57 kDa, 7.4 and 55 deg. C, respectively. Thus, the enzyme was mildly thermophilic as compared to other MerA enzymes. Km and Vmax of the enzyme were 16.9 μM HgCl2 and 6.2 μmol min-1 mg-1 enzyme, respectively. The enzyme was found to be NADPH-specific. To our knowledge this is the first report on characterization of MerA enzyme from an Arthrobacter sp.

  5. Isolation of Indole Utilizing Bacteria Arthrobacter sp. and Alcaligenes sp. From Livestock Waste.

    Science.gov (United States)

    Kim, Minsu; Lee, Jin-Hyung; Kim, Eonmi; Choi, Hyukjae; Kim, Younghoon; Lee, Jintae

    2016-06-01

    Indole is an interspecies and interkingdom signaling molecule widespread in different environmental compartment. Although multifaceted roles of indole in different biological systems have been established, little information is available on the microbial utilization of indole in the context of combating odor emissions from different types of waste. The present study was aimed at identifying novel bacteria capable of utilizing indole as the sole carbon and energy source. From the selective enrichment of swine waste and cattle feces, we identified Gram-positive and Gram-negative bacteria belonging to the genera Arthrobacter and Alcaligenes. Bacteria belonging to the genus Alcaligenes showed higher rates of indole utilization than Arthrobacter. Indole at 1.0 mM for growth was completely utilized by Alcaligenes sp. in 16 h. Both strains produced two intermediates, anthranilic acid and isatin, during aerobic indole metabolism. These isolates were also able to grow on several indole derivatives. Interestingly, an adaptive response in terms of a decrease in cell size was observed in both strains in the presence of indole. The present study will help to explain the degradation of indole by different bacteria and also the pathways through which it is catabolized. Furthermore, these novel bacterial isolates could be potentially useful for the in situ attenuation of odorant indole and its derivatives emitted from different types of livestock waste. PMID:27570307

  6. Genetic differentiation of Arthrobacter population from heavy metal-contaminated environment

    Institute of Scientific and Technical Information of China (English)

    ZHANG Hanbo; REN Weimin; SHAO Qiyong; DUAN Changqun

    2007-01-01

    Six samples containing extremely high concentration of Pb,Zn,and Cd were obtained from the layers of 5-10 cm and 25-30 cm three tailing piles,with ages of about 10,20 and more than 80 years,respectively.Then,48 bacterial strains were obtained from these samples,and subsequently their phylogenetic positions were determined by analysis on the partial sequence of 16S rRNA gene (fragment length ranging from 474 to 708 bp).These isolates were members of the Arthrobacter genus,phylogenetically close to A.keyseri and A.ureafaciens,with sequence ranging from 99.1%to 100%.Furthermore,genetic variation between subpopulations from different samples was revealed by analysis on their randomly amplified polymorphic DNA profile.Nei genetic distance showed that the greatest differentiation occurred between subpopulation A and C.Notably,either genetic distance between subpopulations from the layers of 5-10 cm and 25-30 cm of each tailing pile or between same layers of different tailing pile increased with the history of tailings.Moreover,correlation analysis showed that soluble Pb has a significantly negative relationship with Nei'gene diversity of subpopulation.It was assumed that soluble Pb may be responsible for the reduced genetic diversity of the Arthrobacter population.Our data provided evidence that genetic differentiation of microbial populations was consistent with the changes of environmental factors,particularly heavy metals.

  7. One-pot conversion of levan prepared from Serratia levanicum NN to difructose anhydride IV by Arthrobacter nicotinovorans levan fructotransferase.

    Science.gov (United States)

    Kikuchi, Hiroto; Sakurai, Hiroaki; Nagura, Taizo; Aritsuka, Tsutomu; Tomita, Fusao; Yokota, Atsushi

    2010-03-01

    The newly established difructose anhydride IV (DFA IV) production system is comprised of the effective production of levan from sucrose by Serratia levanicum NN, the conversion of the levan into DFA IV by levan fructotransferase from Arthrobacter nicotinovorans GS-9, which is highly expressed in an Escherichiacoli transformant, and a practical purification step. The chemical properties of DFA IV were also investigated. PMID:20159571

  8. Growth of Arthrobacter sp. strain JBH1 on nitroglycerin as the sole source of carbon and nitrogen.

    Science.gov (United States)

    Husserl, Johana; Spain, Jim C; Hughes, Joseph B

    2010-03-01

    Arthrobacter sp. strain JBH1 was isolated from nitroglycerin-contaminated soil by selective enrichment. Detection of transient intermediates and simultaneous adaptation studies with potential intermediates indicated that the degradation pathway involves the conversion of nitroglycerin to glycerol via 1,2-dinitroglycerin and 1-mononitroglycerin, with concomitant release of nitrite. Glycerol then serves as the source of carbon and energy.

  9. Transcriptional profiling of Gram-positive Arthrobacter in the phyllosphere: induction of pollutant degradation genes by natural plant phenolic compounds

    NARCIS (Netherlands)

    Scheublin, T.R.; Deusch, S.; Moreno-Forero, S.K.; Müller, J.A.; van der Meer, J.R.; Leveau, J.H.J.

    2014-01-01

    Arthrobacter chlorophenolicus A6 is a Gram-positive, 4-chlorophenol degrading soil bacterium that was recently shown to be an effective colonizer of plant leaf surfaces. The genetic basis for this phyllosphere competency is unknown. In this paper, we describe the genome-wide expression profile of A.

  10. Nitrogen metabolism in the facultative methylotroph Arthrobacter P1 grown with various amines or ammonia as nitrogen sources

    NARCIS (Netherlands)

    de Boer, L.; Brouwer, J.W.; Hassel, C.W. van; Levering, Pieter; Dijkhuizen, L.

    1989-01-01

    The metabolism of trimethylamine (TMA) and dimethylamine (DMA) in Arthrobacter P1 involved the enzymes TMA monooxygenase and trimethylamine-N-oxide (TMA-NO) demethylase, and DMA monooxygenase, respectively. The methylamine and formaldehyde produced were further metabolized via a primary amine oxidas

  11. Draft Genome Sequence of a Papaverine-Degrading, Gram-positive Arthrobacter sp., Isolated from Soil Near Hohenheim, Germany

    OpenAIRE

    Reznicek, Ondrej; Facey, Sandra J; Hauer, Bernhard

    2015-01-01

    We present the 4.8-Mb draft genome of a soil bacterium identified as Arthrobacter sp. This Gram-positive soil bacterium is able to use the aromatic compound papaverine as sole carbon source and will be examined for novel oxygenases.

  12. Competition for Ammonium between Plant-Roots and Nitrifying and Heterotrophic Bacteria and the Effects of Protozoan Grazing

    NARCIS (Netherlands)

    Verhagen, F.J.M.; Laanbroek, H.J.; Woldendorp, J.W.

    1995-01-01

    The competition for limiting amounts of ammonium between the chemolithotrophic ammonium-oxidizing species Nitrosomonas europaea, the heterotrophic species Arthrobacter globiformis and roots of Plantago lanceolata (Ribwort plantain) was studied in a series of model systems of increasing complexity, i

  13. Biodegradation of p-nitrophenol via 1,2,4-benzenetriol by an Arthrobacter sp.

    Energy Technology Data Exchange (ETDEWEB)

    Jain, R.K.; Spain, J.C. [Armstrong Lab., Tyndall Air Force Base, FL (United States); Dreisbach, J.H. [Univ. of Scranton, PA (United States)

    1994-08-01

    The degradation of p-nitrophenol (PNP) by Moraxella and Pseudomonas spp. involves an initial monooxygenase-catalyzed removal of the nitro group. The resultant hydroquinone is subject to ring fission catalyzed by a dioxygenase enzyme. A strain of an Arthrobacter sp. JS443, capable of degrading PNP with stoichiometric release of nitrite has been isolated. During induction of the enzymes required for growth on PNP, 1,2,4-benzenetriol was identified as an intermediate by gas chromatography-mass spectroscopy and radiotracer studies. 1,2,4-Benzenetriol was converted to maleylacetic acid, which was further degraded by the beta-ketoadipate pathway. Conversion of PNP to 1,2,4-benzenetriol is catalyzed by a monooxygenase system in strain JS443 through the formation of 4-nitrocatechol, 4-nitroresorcinol, or both. Results clearly indicate the existence of an alternative pathway for the biodegradation of PNP. 15 refs, 2 figs., 2 tabs.

  14. Purification and characterization of alpha-L-arabinofuranosidase from Arthrobacter sp. MTCC 5214 in solid-state fermentation

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Numan, M.T.H.; Mukherjee, B.; Satwekar, A.; Bhosle, N.B.

    protein following the method of bicinchoninic acid with bovine serum albumin as the standard [27]. The protein content of the chromatographic eluents was measured by monitoring the absorbance at 280 nm. 2.6. Preparation of enzyme Erlenmeyer flask (500 ml... cellulose [31] and Aspergillus kawachii [32]. This may indicate the constitutive nature of a-L-AFase production in Arthrobacter sp. 3.2. Growth and enzyme production Aswheatbraninducesoptimalexpressionofa-L-AFaseinSSF,it was used for further studies...

  15. Application of zeolite-Arthrobacter viscosus system for the removal of heavy metal and dye : chromium and azure B

    OpenAIRE

    Rosales, E.; Pazos, M.; Sanromán, M. A.; Tavares, M. T.

    2012-01-01

    A hybrid system combining the ion-exchange properties of a NaY zeolite and the characteristics of the bacterium Arthrobacter viscosus was investigated to treat polluted effluents with dye and toxic metals. In this study, the dye and the metal ion employed were a thiazine dye, Azure B, and chromium (VI), respectively. Initially, the removal of dye by the zeolite was tested. The analysis of dye equilibrium isotherms data was done using Langmuir, Freundlich, Sips and Redlich–Peterson models. Red...

  16. Highly hydrolytic reuteransucrase from probiotic Lactobacillus reuteri strain ATCC 55730

    NARCIS (Netherlands)

    Kralj, S.; Stripling, E.; Sanders, P.; Geel-Schutten, G.H. van; Dijkhuizen, L.

    2005-01-01

    Lactobacillus reuteri strain ATCC 55730 (LB BIO) was isolated as a pure culture from a Reuteri tablet purchased from the BioGaia company. This probiotic strain produces a soluble glucan (reuteran), in which the majority of the linkages are of the α-(1→4) glucosidic type (∼70%). This reuteran also co

  17. Metabolism of amino acid amides in Pseudomonas putida ATCC 12633

    NARCIS (Netherlands)

    Hermes, H.F.M.; Croes, L.M.; Peeters, W.P.H.; Peters, P.J.H.; Dijkhuizen, L.

    1993-01-01

    The metabolism of the natural amino acid L-valine, the unnatural amino acids D-valine, and D-, L-phenylglycine (D-, L-PG), and the unnatural amino acid amides D-, L-phenylglycine amide (D, L-PG-NH2) and L-valine amide (L-Val-NH2) was studied in Pseudomonas putida ATCC 12633. The organism possessed c

  18. 3-Methylindole production is regulated in Clostridium scatologenes ATCC 25775

    Science.gov (United States)

    Aims: 3-Methylindole (3-MI) is a degradation product of L-tryptophan and is both an animal waste malodorant and threat to ruminant health. Culture conditions which influence 3-MI production in Clostridium scatologenes ATCC 25775 were investigated. Methods and Results: Cells cultured in anaerobic ...

  19. Complete Genome Sequence of Mycobacterium vaccae Type Strain ATCC 25954

    KAUST Repository

    Ho, Y. S.

    2012-10-26

    Mycobacterium vaccae is a rapidly growing, nontuberculous Mycobacterium species that is generally not considered a human pathogen and is of major pharmaceutical interest as an immunotherapeutic agent. We report here the annotated genome sequence of the M. vaccae type strain, ATCC 25954.

  20. Complete genome sequence of Campylobacter gracilis ATCC 33236T

    Science.gov (United States)

    The human oral pathogen Campylobacter gracilis has been isolated from periodontal and endodontal infections, and also from non-oral head, neck or lung infections. This study describes the whole-genome sequence of the human periodontal isolate ATCC 33236T (=FDC 1084), which is the first closed genome...

  1. s-triazine degrading bacterial isolate Arthrobacter sp. AK-YN10, a candidate for bioaugmentation of atrazine contaminated soil.

    Science.gov (United States)

    Sagarkar, Sneha; Bhardwaj, Pooja; Storck, Veronika; Devers-Lamrani, Marion; Martin-Laurent, Fabrice; Kapley, Atya

    2016-01-01

    The Arthrobacter sp. strain AK-YN10 is an s-triazine pesticide degrading bacterium isolated from a sugarcane field in Central India with history of repeated atrazine use. AK-YN10 was shown to degrade 99 % of atrazine in 30 h from media supplemented with 1000 mg L(-1) of the herbicide. Draft genome sequencing revealed similarity to pAO1, TC1, and TC2 catabolic plasmids of the Arthrobacter taxon. Plasmid profiling analyses revealed the presence of four catabolic plasmids. The trzN, atzB, and atzC atrazine-degrading genes were located on a plasmid of approximately 113 kb.The flagellar operon found in the AK-YN10 draft genome suggests motility, an interesting trait for a bioremediation agent, and was homologous to that of Arthrobacter chlorophenolicus. The multiple s-triazines degradation property of this isolate makes it a good candidate for bioremediation of soils contaminated by s-triazine pesticides. PMID:26403923

  2. s-triazine degrading bacterial isolate Arthrobacter sp. AK-YN10, a candidate for bioaugmentation of atrazine contaminated soil.

    Science.gov (United States)

    Sagarkar, Sneha; Bhardwaj, Pooja; Storck, Veronika; Devers-Lamrani, Marion; Martin-Laurent, Fabrice; Kapley, Atya

    2016-01-01

    The Arthrobacter sp. strain AK-YN10 is an s-triazine pesticide degrading bacterium isolated from a sugarcane field in Central India with history of repeated atrazine use. AK-YN10 was shown to degrade 99 % of atrazine in 30 h from media supplemented with 1000 mg L(-1) of the herbicide. Draft genome sequencing revealed similarity to pAO1, TC1, and TC2 catabolic plasmids of the Arthrobacter taxon. Plasmid profiling analyses revealed the presence of four catabolic plasmids. The trzN, atzB, and atzC atrazine-degrading genes were located on a plasmid of approximately 113 kb.The flagellar operon found in the AK-YN10 draft genome suggests motility, an interesting trait for a bioremediation agent, and was homologous to that of Arthrobacter chlorophenolicus. The multiple s-triazines degradation property of this isolate makes it a good candidate for bioremediation of soils contaminated by s-triazine pesticides.

  3. Characterization of the in vitro assembly of FtsZ in Arthrobacter strain A3 using light scattering.

    Science.gov (United States)

    Yang, Fan; Zhang, Shanshan; Ding, Shenglong; Hou, Yannan; Yu, Linghui; Chen, Ximing; Xiao, Jianxi

    2016-10-01

    The self-assembly of FtsZ, the bacterial homolog of tubulin, plays an essential role in cell division. Light scattering technique is applied to real-time monitor the in vitro assembly of FtsZ in Arthrobacter strain A3, a newly isolated psychrotrophic bacterium. The critical concentration needed for the assembly is estimated as 6.7μM. The polymerization of FtsZ in Arthrobacter strain A3 requires both GTP and divalent metal ions, while salt is an unfavorable condition for the assembly. The FtsZ polymerizes under a wide range of pHs, with the fastest rate around pH 6.0. The FtsZ from Arthrobacter strain A3 resembles Mycobacterium tuberculosis FtsZ in terms of the dependence on divalent metal ions and the slow polymerization rate, while it is different from M. tuberculosis FtsZ considering the sensitivity to salt and pH. The comparison of FtsZ from different organisms will greatly advance our understanding of the biological role of the key cell division protein. PMID:27164494

  4. A method for the production of D-tagatose using a recombinant Pichia pastoris strain secreting β-D-galactosidase from Arthrobacter chlorophenolicus and a recombinant L-arabinose isomerase from Arthrobacter sp. 22c

    Directory of Open Access Journals (Sweden)

    Wanarska Marta

    2012-08-01

    Full Text Available Abstract Background D-Tagatose is a natural monosaccharide which can be used as a low-calorie sugar substitute in food, beverages and pharmaceutical products. It is also currently being tested as an anti-diabetic and obesity control drug. D-Tagatose is a rare sugar, but it can be manufactured by the chemical or enzymatic isomerization of D-galactose obtained by a β-D-galactosidase-catalyzed hydrolysis of milk sugar lactose and the separation of D-glucose and D-galactose. L-Arabinose isomerases catalyze in vitro the conversion of D-galactose to D-tagatose and are the most promising enzymes for the large-scale production of D-tagatose. Results In this study, the araA gene from psychrotolerant Antarctic bacterium Arthrobacter sp. 22c was isolated, cloned and expressed in Escherichia coli. The active form of recombinant Arthrobacter sp. 22c L-arabinose isomerase consists of six subunits with a combined molecular weight of approximately 335 kDa. The maximum activity of this enzyme towards D-galactose was determined as occurring at 52°C; however, it exhibited over 60% of maximum activity at 30°C. The recombinant Arthrobacter sp. 22c L-arabinose isomerase was optimally active at a broad pH range of 5 to 9. This enzyme is not dependent on divalent metal ions, since it was only marginally activated by Mg2+, Mn2+ or Ca2+ and slightly inhibited by Co2+ or Ni2+. The bioconversion yield of D-galactose to D-tagatose by the purified L-arabinose isomerase reached 30% after 36 h at 50°C. In this study, a recombinant Pichia pastoris yeast strain secreting β-D-galactosidase Arthrobacter chlorophenolicus was also constructed. During cultivation of this strain in a whey permeate, lactose was hydrolyzed and D-glucose was metabolized, whereas D-galactose was accumulated in the medium. Moreover, cultivation of the P. pastoris strain secreting β-D-galactosidase in a whey permeate supplemented with Arthrobacter sp. 22c L-arabinose isomerase resulted in a 90% yield

  5. High-level chromate resistance in Arthrobacter sp. strain FB24 requires previously uncharacterized accessory genes

    Directory of Open Access Journals (Sweden)

    Thompson Dorothea K

    2009-09-01

    Full Text Available Abstract Background The genome of Arthrobacter sp. strain FB24 contains a chromate resistance determinant (CRD, consisting of a cluster of 8 genes located on a 10.6 kb fragment of a 96 kb plasmid. The CRD includes chrA, which encodes a putative chromate efflux protein, and three genes with amino acid similarities to the amino and carboxy termini of ChrB, a putative regulatory protein. There are also three novel genes that have not been previously associated with chromate resistance in other bacteria; they encode an oxidoreductase (most similar to malate:quinone oxidoreductase, a functionally unknown protein with a WD40 repeat domain and a lipoprotein. To delineate the contribution of the CRD genes to the FB24 chromate [Cr(VI] response, we evaluated the growth of mutant strains bearing regions of the CRD and transcript expression levels in response to Cr(VI challenge. Results A chromate-sensitive mutant (strain D11 was generated by curing FB24 of its 96-kb plasmid. Elemental analysis indicated that chromate-exposed cells of strain D11 accumulated three times more chromium than strain FB24. Introduction of the CRD into strain D11 conferred chromate resistance comparable to wild-type levels, whereas deletion of specific regions of the CRD led to decreased resistance. Using real-time reverse transcriptase PCR, we show that expression of each gene within the CRD is specifically induced in response to chromate but not by lead, hydrogen peroxide or arsenate. Higher levels of chrA expression were achieved when the chrB orthologs and the WD40 repeat domain genes were present, suggesting their possible regulatory roles. Conclusion Our findings indicate that chromate resistance in Arthrobacter sp. strain FB24 is due to chromate efflux through the ChrA transport protein. More importantly, new genes have been identified as having significant roles in chromate resistance. Collectively, the functional predictions of these additional genes suggest the

  6. Spectrophotometric evaluation of selenium binding by Saccharomyces cerevisiae ATCC MYA-2200 and Candida utilis ATCC 9950 yeast.

    Science.gov (United States)

    Kieliszek, Marek; Błażejak, Stanisław; Płaczek, Maciej

    2016-05-01

    In this study, the ability of selenium binding the biomas of Saccharomyces cerevisiae ATCC MYA-2200 and Candida utilis ATCC 9950 was investigated. Sodium selenite(IV) salts were added to the experimental media at concentrations of 10, 20, 40, and 60 mg Se(4+) L(-1). In the tested concentration range, one concentration reported a significant reduction in the biomass yield of both yeast strains. Intense growth was observed for C. utilis yeast, which reached the highest biomass yield of 15 gd.w.L(-1) after 24h cultivation in the presence of 10mg Se(4+) L(-1). Based on the use of spectrophotometric method for the determination of selenium content by using Variamine Blue as a chromogenic agent, efficient accumulation of this element in the biomass of the investigated yeast was observed. The highest amount of selenium, that is, 5.64 mg Se(4+)gd.w.(-1), was bound from the environment by S. cerevisiae ATCC MYA-2200 cultured in the presence of 60 mg Se(4+) L(-1) medium 72h Slightly less amount, 5.47 mg Se(4+) gd.w.(-1), was absorbed by C. utilis ATCC 9950 during similar cultural conditions. Based on the results of the biomass yield and the use of selenium from the medium, it can be observed that yeasts of the genus Candida are more efficient in binding this element, and this property finds practical application in the production of selenium-enriched yeast.

  7. Spectrophotometric evaluation of selenium binding by Saccharomyces cerevisiae ATCC MYA-2200 and Candida utilis ATCC 9950 yeast.

    Science.gov (United States)

    Kieliszek, Marek; Błażejak, Stanisław; Płaczek, Maciej

    2016-05-01

    In this study, the ability of selenium binding the biomas of Saccharomyces cerevisiae ATCC MYA-2200 and Candida utilis ATCC 9950 was investigated. Sodium selenite(IV) salts were added to the experimental media at concentrations of 10, 20, 40, and 60 mg Se(4+) L(-1). In the tested concentration range, one concentration reported a significant reduction in the biomass yield of both yeast strains. Intense growth was observed for C. utilis yeast, which reached the highest biomass yield of 15 gd.w.L(-1) after 24h cultivation in the presence of 10mg Se(4+) L(-1). Based on the use of spectrophotometric method for the determination of selenium content by using Variamine Blue as a chromogenic agent, efficient accumulation of this element in the biomass of the investigated yeast was observed. The highest amount of selenium, that is, 5.64 mg Se(4+)gd.w.(-1), was bound from the environment by S. cerevisiae ATCC MYA-2200 cultured in the presence of 60 mg Se(4+) L(-1) medium 72h Slightly less amount, 5.47 mg Se(4+) gd.w.(-1), was absorbed by C. utilis ATCC 9950 during similar cultural conditions. Based on the results of the biomass yield and the use of selenium from the medium, it can be observed that yeasts of the genus Candida are more efficient in binding this element, and this property finds practical application in the production of selenium-enriched yeast. PMID:27049131

  8. Biodegradation of nitroglycerin in porous media and potential for bioaugmentation with Arthrobacter sp. strain JBH1.

    Science.gov (United States)

    Husserl, Johana; Hughes, Joseph B

    2013-07-01

    Nitroglycerin (NG) is a toxic explosive found as a contaminant of soil and groundwater. Several microbial strains are capable of partially reducing the NG molecule to dinitro or mononitroesters. Recently, a strain capable of growing on NG as the sole source of carbon and nitrogen (Arthrobacter sp. strain JBH1) was isolated from contaminated soil. Despite the widespread presence of microbial strains capable of transforming NG in contaminated soils and sediments, the extent of NG biodegradation at contaminated sites is still unknown. In this study column experiments were conducted to investigate the extent of microbial degradation of NG in saturated porous media, specifically after bioaugmentation with JBH1. Initial experiments using sterile, low sorptivity sand, showed mineralization of NG after bioaugmentation with JBH1 in the absence of sources of carbon and nitrogen other than NG. Results could be modeled using a first order degradation rate of 0.14d(-1). Further experiments conducted using contaminated soil with high organic carbon content (highly sorptive) resulted in column effluents that did not contain NG although high dinitroester concentrations were observed. Bioaugmentation with JBH1 in sediments containing strains capable of partial transformation of NG resulted in complete mineralization of NG and faster degradation rates.

  9. Combined bioremediation of atrazine-contaminated soil by Pennisetum and Arthrobacter sp. strain DNS10.

    Science.gov (United States)

    Zhang, Ying; Ge, Shijie; Jiang, Mingyue; Jiang, Zhao; Wang, Zhigang; Ma, Bingbing

    2014-05-01

    Strain DNS10 was isolated from the black soil collected from the northeast of China which had been cultivated with atrazine as the sole nitrogen source. Pennisetum is a common plant in Heilongjiang Province of China. The main objective of this paper was to evaluate the efficiency of plant-microbe joint interactions (Arthrobacter sp. DNS10 + Pennisetum) in atrazine degradation compared with single-strain and single-plant effects. Plant-microbe joint interactions degraded 98.10 % of the atrazine, while single strain and single plant only degraded 87.38 and 66.71 % after a 30-day experimental period, respectively. The results indicated that plant-microbe joint interactions had a better degradation effect. Meanwhile, we found that plant-microbe joint interactions showed a higher microbial diversity. The results of microbial diversity illustrated that the positive effects of cropping could improve soil microbial growth and activity. In addition, we planted atrazine-sensitive plants (soybean) in the soil after repair. The results showed that soybean growth in soil previously treated with the plant-microbe joint interactions treatment was better compared with other treatments after 20 days of growth. This was further proved that the soil is more conducive for crop cultivation. Hence, plant-microbe joint interactions are considered to be a potential tool in the remediation of atrazine-contaminated soil.

  10. Biodegradation of Benzo[a]pyrene by Arthrobacter oxydans B4

    Institute of Scientific and Technical Information of China (English)

    PENG Hui; YIN Hua; DENG Jun; YE Jin-Shao; CHEN Shuo-Na; HE Bao-Yan; ZHANG Na

    2012-01-01

    A bacterial strain,Arthrobacter oxydans (B4),capable of degrading benzo[a]pyrene (BaP) in water body,was isolated from a polycyclic aromatic hydrocarbons-contaminated site.Effects of different factors,such as reaction time,pH value,temperature and organic nutrients,on BaP biodegradation by the strain B4 were studied.After 5 d treatment,the concentration of BaP in mineral salts medium was reduced to 0.318 mg L-1,compared to the initial concentration of 1.000 mg L-1.There was a process of acid formation during the degradation with pH failing from initial 7.01 to 4.61 at 5 d,so keeping the water body under slightly alkaline condition was propitious to BaP degradation.Strain B4 efficiently degraded BaP at 20 to 37 ℃ with addition of organic nutrients.The biodegradation and transformation of BaP mainly occurred on cell surfaces,and extracellular secretions played an important role in these processes.Fourier transform infrared spectroscopy and gas chromatograph-mass spectrometer analyses of metabolites showed that ring cleavage occurred in the BaP degradation process and the resulting metabolically utilizable substrates were generated as sole carbon sources for B4 growth.Furthermore,mineralization extent of metabolites was verified by determining the total organic carbon and inorganic carbon in the degradation system.

  11. The rhizobacterium Arthrobacter agilis produces dimethylhexadecylamine, a compound that inhibits growth of phytopathogenic fungi in vitro.

    Science.gov (United States)

    Velázquez-Becerra, Crisanto; Macías-Rodríguez, Lourdes I; López-Bucio, José; Flores-Cortez, Idolina; Santoyo, Gustavo; Hernández-Soberano, Christian; Valencia-Cantero, Eduardo

    2013-12-01

    Plant diseases caused by fungal pathogens such as Botrytis cinerea and the oomycete Phytophthora cinnamomi affect agricultural production worldwide. Control of these pests can be done by the use of fungicides such as captan, which may have deleterious effects on human health. This study demonstrates that the rhizobacterium Arthrobacter agilis UMCV2 produces volatile organic compounds that inhibit the growth of B. cinerea in vitro. A single compound from the volatile blends, namely dimethylhexadecylamine (DMHDA), could inhibit the growth of both B. cinerea and P. cinnamomi when supplied to the growth medium in low concentrations. DMHDA also inhibited the growth of beneficial fungi Trichoderma virens and Trichoderma atroviride but at much higher concentrations. DMHDA-related aminolipids containing 4, 8, 10, 12, and 14 carbons in the alkyl chain were tested for their inhibitory effect on the growth of the pathogens. The results show that the most active compound from those tested was dimethyldodecylamine. This effect correlates with a decrease in the number of membrane lipids present in the mycelium of the pathogen including eicosanoic acid, (Z)-9-hexadecenoic acid, methyl ester, and (Z)-9-octadecenoic acid, methyl ester. Strawberry leaflets treated with DMHDA were not injured by the compound. These data indicate that DMHDA and related compounds, which can be produced by microorganisms may effectively inhibit the proliferation of certain plant pathogens. PMID:23674267

  12. Plasmid-encoded degradation of p-nitrophenol and 4-nitrocatechol by Arthrobacter protophormiae.

    Science.gov (United States)

    Chauhan, A; Chakraborti, A K; Jain, R K

    2000-04-21

    Arthrobacter protophormiae strain RKJ100 is capable of utilizing p-nitrophenol (PNP) as well as 4-nitrocatechol (NC) as the sole source of carbon, nitrogen and energy. The degradation of PNP and NC by this microorganism takes place through an oxidative route, as stoichiometry of nitrite molecules was observed when the strain was grown on PNP or NC as sole carbon and energy sources. The degradative pathways of PNP and NC were elucidated on the basis of enzyme assays and chemical characterization of the intermediates by TLC, GC, (1)H NMR, GC-MS, UV spectroscopy, and HPLC analyses. Our studies clearly indicate that the degradation of PNP proceeds with the formation of p-benzoquinone (BQ) and hydroquinone (HQ) and is further degraded via the beta-ketoadipate pathway. Degradation of NC involved initial oxidation to generate 1,2,4-benzenetriol (BT) and 2-hydroxy-1,4-benzoquinone; the latter intermediate is then reductively dehydroxylated, forming BQ and HQ, and is further cleaved via beta-ketoadipate to TCA intermediates. It is likely, therefore, that the same set of genes encode the further metabolism of HQ in PNP and NC degradation. A plasmid of approximately 65 kb was found to be responsible for harboring genes for PNP and NC degradation in this strain. This was based on the fact that PNP(-) NC(-) derivatives were devoid of the plasmid and had simultaneously lost their capability to grow at the expense of these nitroaromatic compounds.

  13. Improving stability of a novel dextran-degrading enzyme from marine Arthrobacter oxydans KQ11.

    Science.gov (United States)

    Wang, Delong; Lu, Mingsheng; Wang, Xiaobei; Jiao, Yuliang; Fang, Yaowei; Liu, Zhaopu; Wang, Shujun

    2014-03-15

    Dextranases can hydrolyze dextran, so they are used in the sugar industry to mitigate the milling problems associated with dextran contamination. Few studies have been carried out on the storage stability of dextranase, let alone the dextranase of Arthrobacter oxydans KQ11 isolated from sea mud samples. This study improved the storage stability of dextranase from marine A. oxydans KQ11 by adding enzymatic protective reagents (stabilizer and antiseptic). Initially, the conditions (55 °C and 30 min) for maintaining 50% dextranase activity were obtained. Then, the best stabilizers of dextranase were obtained, namely, glycerol (16%), sodium acetate (18%) and sodium citrate (20%). Results showed that p-hydroxybenzoic acid compound sodium acetate (0.05%), D-sodium isoascorbiate (0.03%), and potassium sorbate (0.05%) were the best antiseptics. Subsequent validation experiment showed that dextranase with enzymatic protective reagents maintained 70.8% and 28.96% activities at the 13th week at 25 and 37 °C, respectively. PMID:24528732

  14. Isolation and characteristics of Arthrobacter sp. strain CW-1 for biodegradation of PAEs

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Isolation of new bacterial strains and recognition of their metabolic activities are highly desirable for sustainability of natural ecosystems. Biodegradation of dimethyl phthalate (DMP) under anoxic conditions has been shown to occur as a series of sequential steps using strain CW-1 isolated from digested sludge of Sibao Wastewater Treatment Plant in Hangzhou, China. The microbial colony on LB medium was yellowish, 3~5 mm in diameter, convex in the center, and embedded in mucous externally.The individual cells of strain CW-1 are irregular rods, measuring (0.6~0.7)×(0.9~1.0) μm, V-shaped, with clubbed ends, Gram positive and without any filaments. 16S rDNA (1438 bp) sequence analysis showed that the strain was related to Arthrobacter sp.CW-1 and can degrade PAEs utilizing nitrate as electron acceptor, but cannot mineralize DMP completely. The degradation pathway was recommended as: dimethyl phthalate (DMP)→monomethyl phthalate (MMP)→phthalic acid (PA). DMP biodegradation was a first order reaction with degradation rate constant of 0.3033 d-1 and half-life 2.25 d. The DMP conversion to PA by CW-1 could be described by using sequential kinetic model.

  15. Adenylate nucleotide levels and energy charge in Arthrobacter crystallopoietes during growth and starvation.

    Science.gov (United States)

    Leps, W T; Ensign, J C

    1979-07-01

    The adenylate nucleotide concentrations, based on internal water space, were determined in cells of Arthrobacter crystallopoietes during growth and starvation and the energy charge of the cells was calculated. The energy charge of spherical cells rose during the first 10 h of growth, then remained nearly constant for as long as 20 h into the stationary phase. The energy charge of rod-shaped cells rose during the first 4 h of growth, then remained constant during subsequent growth and decreased in the stationary growth phase. Both spherical and rod-shaped cells excreted adenosine monophosphate but not adenosine triphosphate or adenosine diphosphate during starvation. The intracellular energy charge of spherical cells declined during the initial 10 h and then remained constant for 1 week of starvation at a value of 0.78. The intracellular energy charge of rod-shaped cells declined during the first 24 h of starvation, remained constant for the next 80 h, then decreased to a value of 0.73 after a total of 168 h starvation. Both cell forms remained more than 90% viable during this time. Addition of a carbon and energy source to starving cells resulted in an increase in the ATP concentration and as a result the energy charge increased to the smae levels as found during growth.

  16. Production of Protocatechuic Acid in Bacillus Thuringiensis ATCC33679

    Directory of Open Access Journals (Sweden)

    Bianca L. Garner

    2012-03-01

    Full Text Available Protocatechuic acid, or 3,4-dihydroxybenzoic acid, is produced by both soil and marine bacteria in the free form and as the iron binding component of the siderophore petrobactin. The soil bacterium, Bacillus thuringiensis kurstaki ATCC 33679, contains the asb operon, but does not produce petrobactin. Iron restriction resulted in diminished B. thuringiensis kurstaki ATCC 33679 growth and the production of catechol(s. The gene product responsible for protocatechuic acid (asbF and its receptor (fatB were expressed during stationary phase growth. Gene expression varied with growth temperature, with optimum levels occurring well below the Bacillus anthracis virulence temperature of 37 °C. Regulation of protocatechuic acid suggests a possible role for this compound during soil growth cycles.

  17. Magnetic response in cultures of Streptococcus mutans ATCC-27607.

    Science.gov (United States)

    Adamkiewicz, V W; Bassous, C; Morency, D; Lorrain, P; Lepage, J L

    1987-01-01

    Streptococcus mutans ATCC-27607 produces exopolysaccharides that adhere to glass. In the normal geomagnetic field about 50% more polysaccharide adhere preferentially to glass surfaces facing North as compared to South facing surfaces. Reversal of the direction of the magnetic field by 180 degrees produces a similar reversal in the direction of the preferential accumulation. Reduction of the field by 90% abolishes the preferential accumulation. PMID:3582582

  18. Genome Sequence of Propionibacterium acnes Type II Strain ATCC 11828

    OpenAIRE

    Horváth, Balázs; Hunyadkürti, Judit; Vörös, Andrea; Fekete, Csaba; Urbán, Edit; Kemény, Lajos; Nagy, István

    2012-01-01

    Propionibacterium acnes is an anaerobic Gram-positive bacterium that forms part of the normal human cutaneous microbiota and is occasionally associated with inflammatory diseases (I. Kurokawa et al., Exp. Dermatol. 18:821–832, 2009). Here we present the complete genome sequence for the commercially available P. acnes type II reference strain ATCC 11828 (I. Nagy et al., Microbes Infect. 8:2195–2205, 2006) recovered from a subcutaneous abscess.

  19. Cellulose produced by Gluconacetobacter xylinus strains ATCC 53524 and ATCC 23768: Pellicle formation, post-synthesis aggregation and fiber density.

    Science.gov (United States)

    Lee, Christopher M; Gu, Jin; Kafle, Kabindra; Catchmark, Jeffrey; Kim, Seong H

    2015-11-20

    The pellicle formation, crystallinity, and bundling of cellulose microfibrils produced by bacterium Gluconacetobacter xylinus were studied. Cellulose pellicles were produced by two strains (ATCC 53524 and ATCC 23769) for 1 and 7 days; pellicles were analyzed with scanning electron microscopy (SEM), X-ray diffraction (XRD), vibrational sum-frequency-generation (SFG) spectroscopy, and attenuated total reflectance infrared (ATR-IR) spectroscopy. The bacterial cell population was higher at the surface exposed to air, indicating that the newly synthesized cellulose is deposited at the top of the pellicle. XRD, ATR-IR, and SFG analyses found no significant changes in the cellulose crystallinity, crystal size or polymorphic distribution with the culture time. However, SEM and SFG analyses revealed cellulose macrofibrils produced for 7 days had a higher packing density at the top of the pellicle, compared to the bottom. These findings suggest that the physical properties of cellulose microfibrils are different locally within the bacterial pellicles. PMID:26344281

  20. Characterizing the Catalytic Potential of Deinococcus, Arthrobacter and other Robust Bacteria in Contaminated Subsurface Environments of the Hanford Site

    Energy Technology Data Exchange (ETDEWEB)

    Daly, Michael J.

    2005-06-01

    Natural selection in highly radioactive waste sites may yield bacteria with favorable bioremediating characteristics. However, until recently the microbial ecology of such environments has remained unexplored because of the high costs and technical complexities associated with extracting and characterizing samples from such sites. We have examined the bacterial ecology within radioactive sediments from a high-level nuclear waste plume in the vadose zone on the DOE?s Hanford Site in south-central Washington state (Fredrickson et al, 2004). Manganese-dependent, radiation resistant bacteria have been isolated from this contaminated site including the highly Mn-dependent Deinococcus and Arthrobacter spp.

  1. Highly hydrolytic reuteransucrase from probiotic Lactobacillus reuteri strain ATCC 55730

    OpenAIRE

    Kralj, S.; Stripling, E.; Sanders, P.; van Geel-Schutten, G.H.; Dijkhuizen, L.

    2005-01-01

    Lactobacillus reuteri strain ATCC 55730 (LB BIO) was isolated as a pure culture from a Reuteri tablet purchased from the BioGaia company. This probiotic strain produces a soluble glucan (reuteran), in which the majority of the linkages are of the α-(1→4) glucosidic type (∼70%). This reuteran also contains α-(1→6)-linked glucosyl units and 4,6-disubstituted α-glucosyl units at the branching points. The LB BIO glucansucrase gene (gtfO) was cloned and expressed in Escherichia coli, and the GTFO ...

  2. Optimization of Endoglucanase Production from a Novel Bacterial Isolate, Arthrobacter sp. HPG166 and Characterization of Its Properties

    Directory of Open Access Journals (Sweden)

    Shengwei Huang

    2015-10-01

    Full Text Available ABSTRACTIn this study, a potential novel cellulolytic bacteriumArthrobacter sp. HPG166 was isolated from the hindgut of root-feeding larvaeHolotrichia parallela. Optimization of fermentation factors for endoglucanase production byArthrobacter sp. HPG166 was carried out via response surface methodology. Sodium carboxymethylcellulose 1.19% (w/v and beef extract 0.35% (w/v were the ideal combination of carbon and nitrogen sources for enzyme production; the optimum temperature and pH for cellulase production were 34°C and pH 8.0 respectively. Under the optimized fermentation conditions, the maximum endoglucanase activity of 1.411 U mL-1 was obtained. The crude endoglucanase was thermotolerant as it retained 50.31% of its activity after incubation at 70°C for an hour. Metal profile of the enzyme indicated that Mg2+ and Na+ were strong stimulators while Mn2+ and Co+ drastically inhibited its activity. Due to its particular characteristics, this enzyme could have potential for industrial applications.

  3. Optimization of four types of antimicrobial agents to increase the inhibitory ability of marine Arthrobacter oxydans KQ11 dextranase mouthwash

    Science.gov (United States)

    Ren, Wei; Wang, Shujun; Lü, Mingsheng; Wang, Xiaobei; Fang, Yaowei; Jiao, Yuliang; Hu, Jianen

    2016-03-01

    We adopted the response surface methodology using single factor and orthogonal experiments to optimize four types of antimicrobial agents that could inhibit biofilm formation by Streptococcus mutans, which is commonly found in the human oral cavity and causes tooth decay. The objective was to improve the function of marine Arthrobacter oxydans KQ11 dextranase mouthwash (designed and developed by our laboratory). The experiment was conducted in a three-level, four-variable central composite design to determine the best combination of ZnSO4, lysozyme, citric acid and chitosan. The optimized antibacterial agents were 2.16 g/L ZnSO4, 14 g/L lysozyme, 4.5 g/L citric acid and 5 g/L chitosan. The biofilm formation inhibition reached 84.49%. In addition, microscopic observation of the biofilm was performed using scanning electron microscopy and confocal laser scanning microscopy. The optimized formula was tested in marine dextranase Arthrobacter oxydans KQ11 mouthwash and enhanced the inhibition of S. mutans. This work may be promoted for the design and development of future marine dextranase oral care products.

  4. 40 CFR 180.1205 - Beauveria bassiana ATCC #74040; exemption from the requirements of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL... the insecticide Beauveria bassiana (ATCC #74040) in or on all food commodities when applied or used...

  5. Arthrobacter P 1, a Fast Growing Versatile Methylotroph with Amine Oxidase as a Key Enzyme in the Metabolism of Methylated Amines

    NARCIS (Netherlands)

    Dijken, J.P. van; Veenhuis, M.; Harder, W.

    1981-01-01

    A facultative methylotrophic bacterium was isolated from enrichment cultures containing methylamine as the sole carbon source. It was tentatively identified as an Arthrobacter species. Extracts of cells grown on methylamine or ethylamine contained high levels of amine oxidase (E.C. 1.4.3) activity.

  6. Regulation of methylamine and formaldehyde metabolism in Arthrobacter P1. Formaldehyde is the inducing signal for the synthesis of the RuMP cycle enzyme hexulose phosphate synthase

    NARCIS (Netherlands)

    Croes, L.M.; Dijkhuizen, L.

    1986-01-01

    The inducing potential of formaldehyde on the synthesis of hexulose phosphate synthase, a key enzyme of the RuMP cycle in Arthrobacter P1, was investigated in resting cell suspensions. Induction of this enzyme only occurred at formaldehyde concentrations of 0.5 mM and below. No evidence was obtained

  7. Production of Biohydrogen from Wastewater by Klebsiella oxytoca ATCC 13182.

    Science.gov (United States)

    Thakur, Veena; Tiwari, K L; Jadhav, S K

    2015-08-01

    Production of biohydrogen from distillery effluent was carried out by using Klebsiella oxytoca ATCC 13182. The work focuses on optimization of pH, temperature, and state of bacteria, which are the various affecting factors for fermentative biohydrogen production. Results indicates that at 35 °C for suspended cultures, the production was at its maximum (i.e., 91.33 ± 0.88 mL) when compared with other temperatures. At 35 °C and at pH 5 and 6, maximum productions of 117.67 ± 1.45 and 111.67 ± 2.72 mL were observed with no significant difference. When immobilized, Klebsiella oxytoca ATCC 13182 was used for biohydrogen production at optimized conditions, production was 186.33 ± 3.17 mL. Hence, immobilized cells were found to be more advantageous for biological hydrogen production over suspended form. Physicochemical analysis of the effluent was conducted before and after fermentation and the values suggested that the fermentative process is an efficient method for biological treatment of wastewater.

  8. Experimental measurements of the radiation characteristics of Anabaena variabilis ATCC 29413-U and Rhodobacter sphaeroides ATCC 49419

    Energy Technology Data Exchange (ETDEWEB)

    Berberoglu, Halil; Pilon, Laurent [Mechanical and Aerospace Engineering Department, Henry Samueli School of Engineering and Applied Science, University of California Los Angeles, Los Angeles, CA 90095 (United States)

    2007-12-15

    The objective of this study is to experimentally measure the radiation characteristics of hydrogen producing microorganisms. Special attention is paid to the filamentous cyanobacteria Anabaena variabilis ATCC 29413-U and the unicellular purple bacteria Rhodobacter sphaeroides ATCC 49419 two of the widely studied photobiological hydrogen producers. The extinction and absorption coefficients are measured in the spectral range from 300 to 1300 nm using a spectrophotometer with and without an integrating sphere. Moreover, a nephelometer has been constructed to measure the scattering phase function of the microorganisms at 632.8 nm. The data are used to recover the mass specific absorption, scattering, and extinction cross-sections, the single scattering albedo, and the scattering phase function of the microorganisms. The scattering phase functions of both microorganisms were peaked strongly in the forward direction as expected from their size parameter and shape. The results reported in this study can be used with the radiative transport equation (RTE) to accurately predict and optimize light transport in photobioreactors for photobiological hydrogen production. Finally, the results show that absorption cross-sections of A. variabilis and R. sphaeroides have peaks that do not overlap but rather enlarge the spectral width of the absorption cross-section of a potential symbiotic culture promising more efficient utilization of solar radiation from light transfer point of view. (author)

  9. Physicochemical factors for affecting the efficiency of Arthrobacter chlorophenolicus MN1409 on manganese oxidation%理化因素对锰细菌Arthrobacter chlorophenolicus MN1409氧化Mn2+的影响

    Institute of Scientific and Technical Information of China (English)

    冯福鑫; 王芳; 许旭萍

    2012-01-01

    Arthrobacter chlorophenolicus MN1409是1株分离自锰矿样品的高效锰氧化细菌.为了建立该菌株氧化Mn2+的适宜条件,研究了接种量、装液量、摇床转速、温度、pH值、Fe2+初始质量浓度和Mn2+初始质量浓度等理化因素对其锰氧化效率的影响.结果表明,接种量、装液量、转速和Mn2+初始质量浓度在一定范围内变化对锰氧化率影响不大;而温度、pH值和Fe2+初始质量浓度变化对锰氧化率有较大影响.当接种量为4%,装液量为60mL,温度为25℃,摇床转速为100r/min,pH值为7,且有一定的Fe2+存在时,Arthrobacter chlorophenolicus MN1409对锰的氧化效率最高.%The physicochemical factors for affecting the manganese oxidation efficiency of an manganese oxidizing bacteria named Arthrobacter chlorophennlicus MN1409 which was isolated from manganese ore sample were studied. The identified factors include inoculum volume, liquid volume, temperature, shaking speed, pH as well as the concentration of Fe2+ and Mn2+ . The results showed that there was no significant effect on the biological manganese oxidation rate and chemical manganese oxidation rate during 2% - 10% inoculum volumes. 4% inoculum volume was the optimum condition according to its highest total manganese oxidation rate. Similarly, biological and chemical manganese oxidation rale have no significant difference for the liquid volume ranging from 50 mL to 100 mL in 250 mL triangular flask , the highest total manganese oxidation rate occurred in 60 mL liquid volume. Nevertheless, temperature showed an positive effect on manganese oxidation rate. By increasing the temperature from 15 ℃ to 35 ℃, highest biological and chemical manganese oxidation rate could be reached at 25 ℃ and 35 ℃ , respectively, plus the total oxidation rate could achieved 81.52% as well in 35 ℃ . In addition, the effect of shaker speed on manganese oxidation efficiency (MOE) showed that the oxidation rates were similar for

  10. Biotransformation of (-)beta-pinene by Aspergillus niger ATCC 9642.

    Science.gov (United States)

    Toniazzo, Geciane; de Oliveira, Débora; Dariva, Cláudio; Oestreicher, Enrique Guillermo; Antunes, Octávio A C

    2005-01-01

    The main objective of this work was to investigate the biotransformations of (-)alpha-pinene, (-)beta-pinene, and (+) limonene by Aspergillus niger ATCC 9642. The culture conditions involved--concentration of cosolvent (EtOH), substrate applied, and sequential addition of substrates were--investigated. Adaptation of the precultures with small amounts of substrate was also studied. The experiments were performed in conical flasks with liquid cultures. This strain of A. niger was able to convert only (-)beta-pinene into alpha-terpineol. An optimum conversion of (-)beta-pinene into alpha-terpineol of about 4% was obtained when the substrate was applied as a diluted solution in EtOH and sequential addition of substrate was used.

  11. Complete genome sequence of Anabaena variabilis ATCC 29413

    Energy Technology Data Exchange (ETDEWEB)

    Thiel, Teresa [University of Missouri, St. Louis; Pratte, Brenda S. [University of Missouri, St. Louis; Zhong, Jinshun [University of Missouri, St. Louis; Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Copeland, A [U.S. Department of Energy, Joint Genome Institute; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Han, Cliff [Los Alamos National Laboratory (LANL); Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute

    2013-01-01

    Anabaena variabilis ATCC 29413 is a filamentous, heterocyst-forming cyanobacterium that has served as a model organism, with an extensive literature extending over 40 years. The strain has three distinct nitrogenases that function under different environmental conditions and is capable of photoautotrophic growth in the light and true heterotrophic growth in the dark using fructose as both carbon and energy source. While this strain was first isolated in 1964 in Mississippi and named Ana-baena flos-aquae MSU A-37, it clusters phylogenetically with cyanobacteria of the genus Nostoc. The strain is a moderate thermophile, growing well at approximately 40 C. Here we provide some additional characteristics of the strain, and an analysis of the complete genome sequence.

  12. Complete Genome Sequencing of Protease-Producing Novel Arthrobacter sp. Strain IHBB 11108 Using PacBio Single-Molecule Real-Time Sequencing Technology

    OpenAIRE

    Kiran, Shashi; Swarnkar, Mohit K.; Pal, Mohinder; Thakur, Rishu; Tewari, Rupinder; Singh, Anil Kumar; Gulati, Arvind

    2015-01-01

    A previously uncharacterized species of the genus Arthrobacter, strain IHBB 11108 (MCC 2780), is a Gram-positive, strictly aerobic, nonmotile, cold-adapted, and protease-producing alkaliphilic actinobacterium, isolated from shallow undersurface water from Chandra Tal Lake, Lahaul-Spiti, India. The complete genome of the strain is 3.6 Mb in size with an average 58.97% G+C content.

  13. Alleviating salt stress in tomato seedlings using Arthrobacter and Bacillus megaterium isolated from the rhizosphere of wild plants grown on saline-alkaline lands.

    Science.gov (United States)

    Fan, Pengfei; Chen, Daitao; He, Yanan; Zhou, Qingxia; Tian, Yongqiang; Gao, Lihong

    2016-11-01

    Salt-induced soil degradation is common in farmlands and limits the growth and development of numerous crop plants in the world. In this study, we isolated salt-tolerant bacteria from the rhizosphere of Tamarix chinensis, Suaeda salsa and Zoysia sinica, which are common wild plants grown on a saline-alkaline land, to test these bacteria's efficiency in alleviating salt stress in tomato plants. We screened out seven strains (TF1-7) that are efficient in reducing salt stress in tomato seedlings. The sequence data of 16S rRNA genes showed that these strains belong to Arthrobacter and Bacillus megaterium. All strains could hydrolyze casein and solubilize phosphate, and showed at least one plant growth promotion (PGP)-related gene, indicating their potential in promoting plant growth. The Arthrobacter strains TF1 and TF7 and the Bacillus megaterium strain TF2 and TF3 could produce indole acetic acid under salt stress, further demonstrating their PGP potential. Tomato seed germination, seedling length, vigor index, and plant fresh and dry weight were enhanced by inoculation of Arthrobacter and B. megaterium strains under salt stress. Our results demonstrated that salt-tolerant bacteria isolated from the rhizosphere of wild plants grown on saline-alkaline lands could be used for alleviating salt stress in crop plants. PMID:27196364

  14. Draft Genome Sequence of Methicillin-Sensitive Staphylococcus aureus ATCC 29213

    OpenAIRE

    Soni, Isha; Chakrapani, Harinath; Chopra, Sidharth

    2015-01-01

    Staphylococcus aureus subsp. aureus ATCC 29213 is one of the most commonly used strains in drug discovery research and for quality control. We report the completed draft genome sequence for the strain.

  15. Colonization and Immunomodulation by Lactobacillus reuteri ATCC 55730 in the Human Gastrointestinal Tract

    OpenAIRE

    Valeur, Nana; Engel, Peter; Carbajal, Noris; Connolly, Eamonn; Ladefoged, Karin

    2004-01-01

    Lactobacillus reuteri ATCC 55730 is a probiotic (health-promoting) bacterium widely used as a dietary supplement. This study was designed to examine local colonization of the human gastrointestinal mucosa after dietary supplementation with L. reuteri ATCC 55730 and to determine subsequent immune responses at the colonized sites. In this open clinical investigation, 10 healthy volunteers and 9 volunteers with ileostomy underwent gastroscopy or ileoscopy and biopsy samples were taken from the s...

  16. Trehalose induces antagonism towards Pythium debaryanum in Pseudomonas fluorescens ATCC 17400.

    OpenAIRE

    Gaballa, A.; Abeysinghe, P D; Urich, G; Matthijs, S.; De Greve, H; Cornelis, P; N. Koedam

    1997-01-01

    Pseudomonas fluorescens ATCC 17400 shows in vitro activity against Pythium debaryanum under conditions of iron limitation. A lacZ reporter gene introduced by transposon mutagenesis into the P. fluorescens ATCC 17400 trehalase gene (treA) was induced by a factor released by the phytopathogen Pythium debaryanum. The induction of the lacZ gene was lost upon treatment of the Pythium supernatant with commercial trehalase. A trehalose concentration as low as 1 microM could induce the expression of ...

  17. Development and evaluation of whole-genome oligonucleotide array for Acidithiobacillus ferrooxidans ATCC 23270

    Institute of Scientific and Technical Information of China (English)

    LI Qian; SHEN Li; LUO Hai-lang; YIN Hua-qun; LIAO Li-qin; QIU Guan-zhou; LIU Xue-duan

    2008-01-01

    To effectively monitor the characteristic of Acidithiobacillus ferrooxidans ATCC 23270 at the whole-genomic level,a whole-genome 50-mer-based oligonucleotide microarray was developed based on the 3 217 ORFs of A.ferrooxidans ATCC 23270 genome.Based on artificial oligonucleotide probes,the results showed that the optimal hybridization temperature was 45 ℃.Specificity tests with the purified PCR amplifications of 5 genes (Sulfide-quinone reductase,Cytochrome C,Iron oxidase,Mercuric resistance protein,Nitrogenase iron protein) of A.ferrooxidans ATCC 23270 indicated that the probes on the arrays appeared to be specific to their corresponding target genes.Based on the WGA hybridization to global transcriptional difference of A.ferrooxidans ATCC 23270 strains cultured with Fe(Ⅱ) and S(0),the developed 50-mer WGA could be used for global transcriptome analysis of A.ferrooxidans ATCC 23270.The detection limit was estimated to be approximately 5 ng with the genomic DNA,and at 100 ng of the DNA concentration,all of the signals reached the saturation.In addition,strong linear relationships were observed between hybridization signal intensity and the target DNA concentrations (r2=0.977 and 0.992).The results indicated that this technology had potential as a specific,sensitive and quantitative tool for detection and identification of the strain A.ferrooxidans ATCC 23270 at the whole-genome level.

  18. Evaluation of Arthrobacter aurescens Strain TC1 as Bioaugmentation Bacterium in Soils Contaminated with the Herbicidal Substance Terbuthylazine.

    Directory of Open Access Journals (Sweden)

    Vera P Silva

    Full Text Available In the last years the chloro-s-triazine active substance terbuthylazine has been increasingly used as an herbicide and may leave residues in the environment which can be of concern. The present study aimed at developing a bioaugmentation tool based on the soil bacterium Arthrobacter aurescens strain TC1 for the remediation of terbuthylazine contaminated soils and at examining its efficacy for both soil and aquatic compartments. First, the feasibility of growing the bioaugmentation bacterium inocula on simple sole nitrogen sources (ammonium and nitrate instead of atrazine, while still maintaining its efficiency to biodegrade terbuthylazine was shown. In sequence, the successful and quick (3 days bioremediation efficacy of ammonium-grown A. aurescens TC1 cells was proven in a natural soil freshly spiked or four-months aged with commercial terbuthylazine at a dose 10× higher than the recommended in corn cultivation, to mimic spill situations. Ecotoxicity assessment of the soil eluates towards a freshwater microalga supported the effectiveness of the bioaugmentation tool. Obtained results highlight the potential to decontaminate soil while minimizing terbuthylazine from reaching aquatic compartments via the soil-water pathway. The usefulness of this bioaugmentation tool to provide rapid environment decontamination is particularly relevant in the event of accidental high herbicide contamination. Its limitations and advantages are discussed.

  19. Production of Xylanase from Arthrobacter sp. MTCC 6915 Using Saw Dust As Substrate under Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Sevanan Murugan

    2011-01-01

    Full Text Available Saw dust was used as substrate for xylanase production from Arthrobacter sp. MTCC 6915. The study of period of incubation, temperature, pH, carbon, and nitrogen sources for xylanase production was optimized. Xylanase production was found to be optimum at an incubation period of 96 hrs (117.0 U/mL, temperature 30°C (105.0 U/mL, and pH 9.0 (102.9 U/mL. The results showed that the xylanase production was found to be higher in the presence of carboxymethylcellulose (176.4 U/mL and dextrose (126.0 U/mL. It was also observed that peptone (170.1 U/mL and beef extract (161.7 U/mL supported maximum xylanase production.The enzyme was characterized and found to be fairly active at pH 9 (764.4 U/mL and temperature 60°C (819 U/mL. Even in the present study, a major difference in the production temperature (30°C and optimal temperature (60°C of the enzyme activity was observed. However, the pH of the production media and the enzyme activity were found to be the same (pH 9.

  20. Evaluation of Arthrobacter aurescens Strain TC1 as Bioaugmentation Bacterium in Soils Contaminated with the Herbicidal Substance Terbuthylazine.

    Science.gov (United States)

    Silva, Vera P; Moreira-Santos, Matilde; Mateus, Carla; Teixeira, Tânia; Ribeiro, Rui; Viegas, Cristina A

    2015-01-01

    In the last years the chloro-s-triazine active substance terbuthylazine has been increasingly used as an herbicide and may leave residues in the environment which can be of concern. The present study aimed at developing a bioaugmentation tool based on the soil bacterium Arthrobacter aurescens strain TC1 for the remediation of terbuthylazine contaminated soils and at examining its efficacy for both soil and aquatic compartments. First, the feasibility of growing the bioaugmentation bacterium inocula on simple sole nitrogen sources (ammonium and nitrate) instead of atrazine, while still maintaining its efficiency to biodegrade terbuthylazine was shown. In sequence, the successful and quick (3 days) bioremediation efficacy of ammonium-grown A. aurescens TC1 cells was proven in a natural soil freshly spiked or four-months aged with commercial terbuthylazine at a dose 10× higher than the recommended in corn cultivation, to mimic spill situations. Ecotoxicity assessment of the soil eluates towards a freshwater microalga supported the effectiveness of the bioaugmentation tool. Obtained results highlight the potential to decontaminate soil while minimizing terbuthylazine from reaching aquatic compartments via the soil-water pathway. The usefulness of this bioaugmentation tool to provide rapid environment decontamination is particularly relevant in the event of accidental high herbicide contamination. Its limitations and advantages are discussed. PMID:26662024

  1. TIME DEPENDENT ACCUMULATION OF NICOTINE DERIVATIVES IN THE CULTURE MEDIUM OF ARTHROBACTER NICOTINOVORANS pAO1

    Directory of Open Access Journals (Sweden)

    Răzvan Ștefan Boiangiu

    2014-12-01

    Full Text Available Previous studies have shown that the metabolic intermediate 6-hidroxy-D-nicotine (6HNic found in the Arthrobacter nicotinovorans pAO1+ nicotine catabolic pathway has the ability to bind nicotinic acetylcholine receptors and to sustain spatial memory in rats. These properties make 6HNic a valuable compound with some potential for medical applications, thereby a suitable, simple and efficient method for producing 6-hidroxy-D-nicotine is necessary. Here, we focus on identifying the best moment for harvesting A. nicotinovorans cells in order to directly convert nicotine to 6HNic with the best yield.  The growth of  A. nicotinovorans pAO1+ was monitored and the correlation between the growth phases and nicotine metabolism was established. After about 5 hours of lag,the strain entered the log phase and was fully grown after 10 hours. The nicotine concentration began to drop dramatically as the pAO1+ culture reached saturation and was depleted in 5 hours. As the nicotine concentration dropped, 6HNic began to accumulate, reaching the maximum levels after about 11 hours of growth. Two other products could be detected by HPLC, one which was identified as the nicotine-blue (NB pigment and a second a still unknown end-product. 

  2. Evaluation of 4-bromophenol biodegradation in mixed pollutants system by Arthrobacter chlorophenolicus A6 in an upflow packed bed reactor.

    Science.gov (United States)

    Sahoo, Naresh Kumar; Pakshirajan, Kannan; Ghosh, Pranab Kumar

    2014-09-01

    Bromophenol is listed as priority pollutant by U.S. EPA, however, there is no report so far on its removal in mixed pollutants system by any biological reactor operated in continuous mode. Furthermore, bromophenol along with chlorophenol and nitrophenol are usually the major constituents of paper pulp and pesticide industrial effluent. The present study investigated simultaneous biodegradation of these three pollutants with specially emphasis on substrate competition and crossed inhibition by Arthrobacter chlorophenolicus A6 in an upflow packed bed reactor (UPBR). A 2(3) full factorial design was employed with these pollutants at two different levels by varying their influent concentration in the range of 250-450 mg l(-1). Almost complete removal of all these pollutants and 97 % effluent toxicity removal were achieved in the UPBR at a pollutant loading rate of 1707 mg l(-1) day(-1) or lesser. However, at higher loading rates, the reactor performance deteriorated due to transient accumulation of toxic intermediates. Statistical analysis of the results revealed a strong negative interaction of 4-CP on 4-NP biodegradation. On the other hand, interaction effect between 4-CP and 4-BP was found to be insignificant. Among these three pollutants 4-NP preferentially degraded, however, 4-CP exerted more inhibitory effect on 4-NP biodegradation. This study demonstrated the potential of A. chlorophenolicus A6 for biodegradation of 4-BP in mixed pollutants system by a flow through UPBR system. PMID:24934870

  3. Transcriptional profiling of Gram-positive Arthrobacter in the phyllosphere: induction of pollutant degradation genes by natural plant phenolic compounds.

    Science.gov (United States)

    Scheublin, Tanja R; Deusch, Simon; Moreno-Forero, Silvia K; Müller, Jochen A; van der Meer, Jan Roelof; Leveau, Johan H J

    2014-07-01

    Arthrobacter chlorophenolicus A6 is a Gram-positive, 4-chlorophenol-degrading soil bacterium that was recently shown to be an effective colonizer of plant leaf surfaces. The genetic basis for this phyllosphere competency is unknown. In this paper, we describe the genome-wide expression profile of A.chlorophenolicus on leaves of common bean (Phaseolus vulgaris) compared with growth on agar surfaces. In phyllosphere-grown cells, we found elevated expression of several genes known to contribute to epiphytic fitness, for example those involved in nutrient acquisition, attachment, stress response and horizontal gene transfer. A surprising result was the leaf-induced expression of a subset of the so-called cph genes for the degradation of 4-chlorophenol. This subset encodes the conversion of the phenolic compound hydroquinone to 3-oxoadipate, and was shown to be induced not only by 4-chlorophenol but also hydroquinone, its glycosylated derivative arbutin, and phenol. Small amounts of hydroquinone, but not arbutin or phenol, were detected in leaf surface washes of P.vulgaris by gas chromatography-mass spectrometry. Our findings illustrate the utility of genomics approaches for exploration and improved understanding of a microbial habitat. Also, they highlight the potential for phyllosphere-based priming of bacteria to stimulate pollutant degradation, which holds promise for the application of phylloremediation. PMID:24373130

  4. Fermentation of residual glycerol by Clostridium acetobutylicum ATCC 824 in pure and mixed cultures.

    Science.gov (United States)

    Dams, Rosemeri I; Guilherme, Alexandre A; Vale, Maria S; Nunes, Vanja F; Leitão, Renato C; Santaella, Sandra T

    2016-12-01

    The aim of this research was to estimate the production of hydrogen, organic acids and alcohols by the strain of Clostridium acetobutylicum ATCC 824 using residual glycerol as a carbon source. The experiments were carried out in pure and mixed cultures in batch experiments. Three different sources of inocula for mixed culture were used. Ruminal liquid from goats and sludge collected from two upflow anaerobic sludge blanket reactors treating municipal wastewater and brewery effluent were tested for hydrogen, organic acids and alcohols production with or without C. acetobutylicum ATCC 824. The main detected end-products from the glycerol fermentation were hydrogen, organic acids (acetic, propionic, butyric and caproic) and alcohol (ethanol and 1,3-propanediol - 1,3PD). High hydrogen (0.44 mol H2/mol glycerol consumed) and 1,3PD (0.32 mol 1,3PD/mol glycerol consumed) yields were obtained when the strain C. acetobutylicum ATCC 824 was bioaugmented into the sludge from municipal wastewater using 5 g/L of glycerol. Significant concentrations of n-caproic acid were detected in the ruminal liquid when amended with C. acetobutylicum ATCC 824. The results suggest that glycerol can be used for the generation of H2, 1,3PD and n-caproic acid using C. acetobutylicum ATCC 824 as agent in pure or mixed cultures.

  5. Isolation and characterization of atrazine-degrading Arthrobacter sp. AD26 and use of this strain in bioremediation of contaminated soil

    Institute of Scientific and Technical Information of China (English)

    LI Qingyan; LI Ying; ZHU Xikun; CAI Baoli

    2008-01-01

    A bacterial strain (AD26) capable of utilizing atrazine as a sole nitrogen source for growth was isolated from an industrial wastewatersample by enrichment culture. The 16S rRNA gene sequencing identified AD26 as anArthrobacter sp. PCR assays indicated that AD26contained atrazine-degrading genes trzN and atzBC. The trzN gene of AD26 only differs from the trzN ofArthrobacter aurescens TC1by one base (A→T at 907) and one amino acid (Met→Leu at 303). The specific activity of trzN of AD26 in crude cell extract was0.28 U/mg, which was 1.2 times that of TC 1. This strain has shown faster growth and atrazine-degradation rates in atrazine-containingminimal media than two well characterized atrazine-degrading bacteria, Pseudomonas sp. ADP and Arthrobacter aurescens TC 1. Afterincubating for 48 h at 30℃, the OD600 of AD26 reached 2.6 compared with 1.33 of ADP. AD26 was capable of degrading 500 mg/Lof atrazine in minimal medium at 95% in 72 h, while the degradative rates by TC1 and ADP were only 90% and 86%, respectively. Abioremediation trial of contaminated soil has indicated that AD26 can degrade as high as 98% of atrazine contained in soil (300 mg/kg)after incubating for 20 d at 26℃, nominating this strain as a good candidate for use in bioremediation programs.

  6. Enhanced U(VI) release from autunite mineral by aerobic Arthrobacter sp. in the presence of aqueous bicarbonate

    Energy Technology Data Exchange (ETDEWEB)

    Katsenovich, Yelena; Carvajal, Denny A.; Wellman, Dawn M.; Lagos, Leonel

    2012-04-20

    The bacterial effect on U(VI) leaching from the autunite mineral (Ca[(UO{sub 2})(PO{sub 4})]{sub 2} {center_dot} 3H{sub 2}O) was investigated to provide a more comprehensive understanding into important microbiological processes affecting autunite stability within subsurface bicarbonate-bearing environments. Experiments were performed in a culture of G975 Arthrobacter oxydans strain, herein referred to as G975, a soil bacterium previously isolated from Hanford Site soil. 91 mg of autunite powder and 50 mL of phosphorus-limiting sterile media were amended with bicarbonate ranging between 1-10 mM in glass reactor bottles and inoculated with G975 strain after the dissolution of autunite was at steady state. SEM observations indicated G975 formed a biofilm on the autunite surface and penetrated the mineral cleavages. The mineral surface colonization by bacteria tended to increase concomitantly with bicarbonate concentrations. Additionally, a sterile cultureware with inserts was used in non-contact bioleaching experiments where autunite and bacteria cells were kept separately. The data suggest the G975 bacteria is able to enhance U(VI) leaching from autunite without the direct contact with the mineral. In the presence of bicarbonate, the damage to bacterial cells caused by U(VI) toxicity was reduced, yielding similar values for total organic carbon (TOC) degradation and cell density compared to U(VI)-free controls. The presence of active bacterial cells greatly enhanced the U(VI) bioleaching from autunite in bicarbonate-amended media.

  7. Enhanced U(VI) release from autunite mineral by aerobic Arthrobacter sp. in the presence of aqueous bicarbonate

    Energy Technology Data Exchange (ETDEWEB)

    Katsenovich, Yelena P.; Carvajal, Denny A.; Wellman, Dawn M.; Lagos, Leonel E.

    2012-05-01

    The bacterial effect on U(VI) release from the autunite mineral (Ca[(UO2)(PO4)]2•3H2O) was investigated to provide a more comprehensive understanding of the important microbiological processes affecting autunite stability within subsurface bicarbonate-bearing environments. Experiments were performed in a culture of the Arthrobacter oxydans G975 strain, herein referred to as G975, a soil bacterium previously isolated from Hanford Site soil. 91 mg of autunite powder and 50 mL of phosphorous-limiting sterile media were amended with bicarbonate (ranging between 1 and 10 mM) in glass reactor bottles and inoculated with the G975 strain after the dissolution of autunite was at steady state. SEM observations indicated that G975 formed a biofilm on the autunite surface and penetrated the mineral cleavages. The mineral surface colonization by bacteria tended to increase concomitantly with bicarbonate concentrations. Additionally, a sterile culture-ware with inserts was used in non-contact dissolution experiments where autunite and bacteria cells were kept separately. The data suggest that G975 bacteria is able to enhance the release of U(VI) from autunite without direct contact with the mineral. In the presence of bicarbonate, the damage to bacterial cells caused by U(VI) toxicity was reduced, yielding similar values for total organic carbon (TOC) degradation and cell density compared to U(VI)-free controls. The presence of active bacterial cells greatly enhanced the release of U(VI) from autunite in bicarbonate-amended media.

  8. Proizvodnja β-fruktofuranozidaze s pomoću bakterije Arthrobacter sp. i primjena tog enzima u pretvorbi steviozida i rebaudiozida A

    OpenAIRE

    Xu, Zhong-Wei; Li, Yu-Qiang; Wang, Yong-Hua; Yang, Bo; Ning, Zheng-Xiang

    2009-01-01

    U proizvodnji enzima β-fruktofuranozidaze upotrijebljen je soj bakterije Arthrobacter sp. 10137. Kao najbolji izvori ugljika i dušika za proizvodnju enzima u pokusu na tresilici upotrijebljeni su saharoza i kukuruzni ekstrakt u omjeru 10:1. Maksimalna aktivnost β-fruktofuranozidaze od 26,69 U/mL postignuta je šaržnim uzgojem nakon 22,5 h. Sirovi enzim β-fruktofuranozidaza, dobiven ultrafiltracijom i frakcioniranjem s (NH4)2SO4, pročišćen je 7 puta, što je potvrđeno usporedbom specifične aktiv...

  9. Draft Genome Sequences of Sanguibacteroides justesenii, gen. nov., sp. nov., Strains OUH 308042T (= ATCC BAA-2681T) and OUH 334697 (= ATCC BAA-2682), Isolated from Blood Cultures from Two Different Patients

    DEFF Research Database (Denmark)

    Sydenham, Thomas Vognbjerg; Hasman, Henrik; Justesen, Ulrik Stenz

    2015-01-01

    We announce here the draft genome sequences of Sanguibacteroides justesenii, gen. nov., sp. nov., strains OUH 308042T (= DSM 28342T = ATCC BAA-2681T) and OUH 334697 (= DSM 28341 = ATCC BAA-2682), isolated from blood cultures from two different patients and composed of 51 and 39 contigs for totals...

  10. Draft Genome Sequence of the Microbispora sp. Strain ATCC-PTA-5024, Producing the Lantibiotic NAI-107

    DEFF Research Database (Denmark)

    Sosio, M.; Gallo, G.; Pozzi, R.;

    2014-01-01

    We report the draft genome sequence of Microbispora sp. strain ATCC-PTA-5024, a soil isolate that produces NAI-107, a new lantibiotic with the potential to treat life-threatening infections caused by multidrug-resistant Gram-positive pathogens. The draft genome of strain Microbispora sp. ATCC...

  11. Genome sequence and plasmid transformation of the model high-yield bacterial cellulose producer Gluconacetobacter hansenii ATCC 53582

    Science.gov (United States)

    Florea, Michael; Reeve, Benjamin; Abbott, James; Freemont, Paul S.; Ellis, Tom

    2016-03-01

    Bacterial cellulose is a strong, highly pure form of cellulose that is used in a range of applications in industry, consumer goods and medicine. Gluconacetobacter hansenii ATCC 53582 is one of the highest reported bacterial cellulose producing strains and has been used as a model organism in numerous studies of bacterial cellulose production and studies aiming to increased cellulose productivity. Here we present a high-quality draft genome sequence for G. hansenii ATCC 53582 and find that in addition to the previously described cellulose synthase operon, ATCC 53582 contains two additional cellulose synthase operons and several previously undescribed genes associated with cellulose production. In parallel, we also develop optimized protocols and identify plasmid backbones suitable for transformation of ATCC 53582, albeit with low efficiencies. Together, these results provide important information for further studies into cellulose synthesis and for future studies aiming to genetically engineer G. hansenii ATCC 53582 for increased cellulose productivity.

  12. Characterization of an exo-inulinase from Arthrobacter: a novel NaCl-tolerant exo-inulinase with high molecular mass.

    Science.gov (United States)

    Shen, Jidong; Zhang, Rui; Li, Junjun; Tang, Xianghua; Li, Ruixian; Wang, Min; Huang, Zunxi; Zhou, Junpei

    2015-01-01

    A glycoside hydrolase family 32 exo-inulinase gene was cloned from Arthrobacter sp. HJ7 isolated from saline soil located in Heijing town. The gene encodes an 892-residue polypeptide with a calculated mass of 95.1 kDa and a high total frequency of amino acid residues G, A, and V (30.0%). Escherichia coli BL21 (DE3) cells were used as hosts to express the exo-inulinase gene. The recombinant exo-inulinase (rInuAHJ7) showed an apparently maximal activity at pH 5.0-5.5 and 40-45°C. The addition of 1.0 and 10.0 mM Zn(2+) and Pb(2+) had little or no effect on the enzyme activity. rInuAHJ7 exhibited good salt tolerance, retaining more than 98% inulinase activity at a concentration of 3.0%-20.0% (w/v) NaCl. Fructose was the main product of inulin, levan, and Jerusalem artichoke tubers hydrolyzed by the enzyme. The present study is the first to report the identification and characterization of an Arthrobacter sp exo-inulinase showing a high molecular mass of 95.1 kDa and NaCl tolerance. These results suggest that the exo-inulinase might be an alternative material for potential applications in processing seafood and other foods with high saline contents, such as marine algae, pickles, and sauces. PMID:25695343

  13. Key enzymes enabling the growth of Arthrobacter sp. strain JBH1 with nitroglycerin as the sole source of carbon and nitrogen.

    Science.gov (United States)

    Husserl, Johana; Hughes, Joseph B; Spain, Jim C

    2012-05-01

    Flavoprotein reductases that catalyze the transformation of nitroglycerin (NG) to dinitro- or mononitroglycerols enable bacteria containing such enzymes to use NG as the nitrogen source. The inability to use the resulting mononitroglycerols limits most strains to incomplete denitration of NG. Recently, Arthrobacter strain JBH1 was isolated for the ability to grow on NG as the sole source of carbon and nitrogen, but the enzymes and mechanisms involved were not established. Here, the enzymes that enable the Arthrobacter strain to incorporate NG into a productive pathway were identified. Enzyme assays indicated that the transformation of nitroglycerin to mononitroglycerol is NADPH dependent and that the subsequent transformation of mononitroglycerol is ATP dependent. Cloning and heterologous expression revealed that a flavoprotein catalyzes selective denitration of NG to 1-mononitroglycerol (1-MNG) and that 1-MNG is transformed to 1-nitro-3-phosphoglycerol by a glycerol kinase homolog. Phosphorylation of the nitroester intermediate enables the subsequent denitration of 1-MNG in a productive pathway that supports the growth of the isolate and mineralization of NG.

  14. Identification of a recombinant inulin fructotransferase (difructose dianhydride III forming) from Arthrobacter sp. 161MFSha2.1 with high specific activity and remarkable thermostability.

    Science.gov (United States)

    Wang, Xiao; Yu, Shuhuai; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

    2015-04-01

    Difructose dianhydride III (DFA III) is a functional carbohydrate produced from inulin by inulin fructotransferase (IFTase, EC 4.2.2.18). In this work, an IFTase gene from Arthrobacter sp. 161MFSha2.1 was cloned and expressed in Escherachia coli. The recombinant enzyme was purified by metal affinity chromatography. It showed significant inulin hydrolysis activity, and the produced main product from inulin was determined as DFA III by nuclear magnetic resonance analysis. The molecular mass of the purified protein was calculated to be 43 and 125 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively, suggesting the native enzyme might be a homotrimer. The recombinant enzyme showed maximal activity as 2391 units/mg at pH 6.5 and 55 °C. It displayed the highest thermostability among previously reported IFTases (DFA III forming) and was stable up to 80 °C for 4 h of incubation. The smallest substrate was determined as nystose. The conversion ratio of inulin to DFA III reached 81% when 100 g/L inulin was catalyzed by 80 nM recombinant enzyme for 20 min at pH 6.5 and 55 °C. All of these data indicated that the IFTase (DFA III forming) from Arthrobacter sp. 161MFSha2.1 had great potential for industrial DFA III production.

  15. Evaluation of chromate reductase activity in the cell-free culture filtrate of Arthrobacter sp. SUK 1201 isolated from chromite mine overburden.

    Science.gov (United States)

    Dey, Satarupa; Paul, A K

    2016-08-01

    Arthrobacter sp. SUK 1201, a chromate resistant and reducing bacterium isolated from chromite mine overburden of Sukinda valley, Odisha, India has been evaluated for its hexavalent chromium [Cr(VI)] reduction potential using cell-free culture filtrate as extracellular chromate reductase enzyme. Production of the enzyme was enhanced in presence of Cr(VI) and its reducing efficiency was increased with increasing concentration of Cr(VI). The Michaelis-Menten constant (Km) and the maximum specific velocity (Vmax) of the extracellular Cr(VI) reductase were calculated to be 54.03 μM Cr(VI) and 5.803 U mg(-1) of protein respectively showing high affinity towards Cr(VI). The reducing activity of the enzyme was maximum at pH 6.5-7.5 and at a temperature of 35 °C and was dependent on NADH. The enzyme was tolerant to different metals such as Mn(II), Mg(II) and Fe(III) and was able to reduce Cr(VI) present in chromite mine seepage. These findings suggest that the extracellular chromate reductase of Arthrobacter sp. SUK 1201 has a great promise for use in Cr(VI) detoxification under different environmental conditions, particularly in the mining waste water treatment systems. PMID:27176938

  16. Genome sequence of the vertebrate gut symbiont Lactobacillus reuteri ATCC 53608.

    Science.gov (United States)

    Heavens, Darren; Tailford, Louise E; Crossman, Lisa; Jeffers, Faye; Mackenzie, Donald A; Caccamo, Mario; Juge, Nathalie

    2011-08-01

    Lactobacillus reuteri, inhabiting the gastrointestinal tracts of a range of vertebrates, is a true symbiont with effects established as beneficial to the host. Here we describe the draft genome of L. reuteri ATCC 53608, isolated from a pig. The genome sequence provides important insights into the evolutionary changes underlying host specialization. PMID:21622738

  17. Genome Sequence of the Vertebrate Gut Symbiont Lactobacillus reuteri ATCC 53608 ▿

    OpenAIRE

    Heavens, Darren; Tailford, Louise E.; Crossman, Lisa; Jeffers, Faye; MacKenzie, Donald A.; Caccamo, Mario; Juge, Nathalie

    2011-01-01

    Lactobacillus reuteri, inhabiting the gastrointestinal tracts of a range of vertebrates, is a true symbiont with effects established as beneficial to the host. Here we describe the draft genome of L. reuteri ATCC 53608, isolated from a pig. The genome sequence provides important insights into the evolutionary changes underlying host specialization.

  18. Complete Genome Sequence of Thiostrepton-Producing Streptomyces laurentii ATCC 31255.

    Science.gov (United States)

    Doi, Katsumi; Fujino, Yasuhiro; Nagayoshi, Yuko; Ohshima, Toshihisa; Ogata, Seiya

    2016-06-02

    Streptomyces laurentii ATCC 31255 produces thiostrepton, a thiopeptide class antibiotic. Here, we report the complete genome sequence for this strain, which contains a total of 8,032,664 bp, 7,452 predicted coding sequences, and a G+C content of 72.3%.

  19. Phenotypic and Transcriptomic Analyses of Mildly and Severely Salt-Stressed Bacillus cereus ATCC 14579 Cells

    NARCIS (Netherlands)

    Besten, den H.M.W.; Mols, J.M.; Moezelaar, R.; Zwietering, M.H.; Abee, T.

    2009-01-01

    Bacteria are able to cope with the challenges of a sudden increase in salinity by activating adaptation mechanisms. In this study, exponentially growing cells of the pathogen Bacillus cereus ATCC 14579 were exposed to both mild (2.5% [wt/vol] NaCl) and severe (5% [wt/vol] NaCl) salt stress condition

  20. Complete Genome Sequence of the Quality Control Strain Staphylococcus aureus subsp. aureus ATCC 25923.

    Science.gov (United States)

    Treangen, Todd J; Maybank, Rosslyn A; Enke, Sana; Friss, Mary Beth; Diviak, Lynn F; Karaolis, David K R; Koren, Sergey; Ondov, Brian; Phillippy, Adam M; Bergman, Nicholas H; Rosovitz, M J

    2014-01-01

    Staphylococcus aureus subsp. aureus ATCC 25923 is commonly used as a control strain for susceptibility testing to antibiotics and as a quality control strain for commercial products. We present the completed genome sequence for the strain, consisting of the chromosome and a 27.5-kb plasmid. PMID:25377701

  1. Complete Genome Sequence of the Quality Control Strain Staphylococcus aureus subsp. aureus ATCC 25923

    OpenAIRE

    Todd J. Treangen; Maybank, Rosslyn A.; Enke, Sana; Friss, Mary Beth; Diviak, Lynn F.; Karaolis, David K. R.; Koren, Sergey; Ondov, Brian; Phillippy, Adam M.; Bergman, Nicholas H.; Rosovitz, M. J.

    2014-01-01

    Staphylococcus aureus subsp. aureus ATCC 25923 is commonly used as a control strain for susceptibility testing to antibiotics and as a quality control strain for commercial products. We present the completed genome sequence for the strain, consisting of the chromosome and a 27.5-kb plasmid.

  2. Complete Genome Sequence of the Larval Shellfish Pathogen Vibrio tubiashii Type Strain ATCC 19109.

    Science.gov (United States)

    Richards, Gary P; Needleman, David S; Watson, Michael A; Bono, James L

    2014-12-18

    Vibrio tubiashii is a larval shellfish pathogen. Here, we report the first closed genome sequence for this species (ATCC type strain 19109), which consists of two chromosomes (3,294,490 and 1,766,582 bp), two megaplasmids (251,408 and 122,808 bp), and two plasmids (57,076 and 47,973 bp).

  3. 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

  4. Ca2+-Citrate Uptake and Metabolism in Lactobacillus casei ATCC 334

    NARCIS (Netherlands)

    Mortera, Pablo; Pudlik, Agata; Magni, Christian; Alarcon, Sergio; Lolkema, Juke S.

    2013-01-01

    The putative citrate metabolic pathway in Lactobacillus casei ATCC 334 consists of the transporter CitH, a proton symporter of the citrate-divalent metal ion family of transporters CitMHS, citrate lyase, and the membrane-bound oxaloacetate decarboxylase complex OAD-ABDH. Resting cells of Lactobacill

  5. Complete Genome Sequence of Thiostrepton-Producing Streptomyces laurentii ATCC 31255

    Science.gov (United States)

    Fujino, Yasuhiro; Nagayoshi, Yuko; Ohshima, Toshihisa; Ogata, Seiya

    2016-01-01

    Streptomyces laurentii ATCC 31255 produces thiostrepton, a thiopeptide class antibiotic. Here, we report the complete genome sequence for this strain, which contains a total of 8,032,664 bp, 7,452 predicted coding sequences, and a G+C content of 72.3%. PMID:27257211

  6. Complete Genome Sequence of Streptococcus equi subsp. zooepidemicus Strain ATCC 35246

    OpenAIRE

    Ma, Zhe; Geng, Jianing; Zhang, Hui; Yu, Haiying; Yi, Li; Lei, Meng; Lu, Cheng-Ping; Fan, Hong-Jie; Hu, Songnian

    2011-01-01

    Streptococcus equi subsp. zooepidemicus is an opportunistic pathogen. It has caused a very large economic loss in the swine industry of China and has become a threat to human health. We announce the complete genome sequence of S. equi subsp. zooepidemicus strain ATCC 35246, which provides opportunities to understand its pathogenesis mechanism and genetic basis.

  7. Complete genome sequence of Streptococcus equi subsp. zooepidemicus strain ATCC 35246.

    Science.gov (United States)

    Ma, Zhe; Geng, Jianing; Zhang, Hui; Yu, Haiying; Yi, Li; Lei, Meng; Lu, Cheng-ping; Fan, Hong-jie; Hu, Songnian

    2011-10-01

    Streptococcus equi subsp. zooepidemicus is an opportunistic pathogen. It has caused a very large economic loss in the swine industry of China and has become a threat to human health. We announce the complete genome sequence of S. equi subsp. zooepidemicus strain ATCC 35246, which provides opportunities to understand its pathogenesis mechanism and genetic basis. PMID:21914890

  8. Interaction of Pseudomonas putida ATCC 12633 and Bacteriophage gh-1 in Berea Sandstone Rock.

    Science.gov (United States)

    Chang, P L; Yen, T F

    1985-12-01

    Measurements of the passage of Pseudomonas putida ATCC 12633 and a phage-resistant mutant through Berea sandstone rock were made. When bacteriophage gh-1 was adsorbed within the rock matrix, a reduction in the passage of the susceptible but not the resistant cells through the rock was observed.

  9. Genome sequence of the Bacteroides fragilis phage ATCC 51477-B1

    OpenAIRE

    Hawkins Shawn A; Layton Alice C; Ripp Steven; Williams Dan; Sayler Gary S

    2008-01-01

    Abstract The genome of a fecal pollution indicator phage, Bacteroides fragilis ATCC 51477-B1, was sequenced and consisted of 44,929 bases with a G+C content of 38.7%. Forty-six putative open reading frames were identified and genes were organized into functional clusters for host specificity, lysis, replication and regulation, and packaging and structural proteins.

  10. Genome sequence of the Bacteroides fragilis phage ATCC 51477-B1

    OpenAIRE

    Hawkins, Shawn A; Layton, Alice C.; Ripp, Steven; Williams, Dan; Sayler, Gary S.

    2008-01-01

    The genome of a fecal pollution indicator phage, Bacteroides fragilis ATCC 51477-B1, was sequenced and consisted of 44,929 bases with a G+C content of 38.7%. Forty-six putative open reading frames were identified and genes were organized into functional clusters for host specificity, lysis, replication and regulation, and packaging and structural proteins.

  11. Low temperature reduction of hexavalent chromium by a microbial enrichment consortium and a novel strain of Arthrobacter aurescens

    Directory of Open Access Journals (Sweden)

    Thompson Vicki S

    2006-01-01

    Full Text Available Abstract Background Chromium is a transition metal most commonly found in the environment in its trivalent [Cr(III] and hexavalent [Cr(VI] forms. The EPA maximum total chromium contaminant level for drinking water is 0.1 mg/l (0.1 ppm. Many water sources, especially underground sources, are at low temperatures (less than or equal to 15 Centigrade year round. It is important to evaluate the possibility of microbial remediation of Cr(VI contamination using microorganisms adapted to these low temperatures (psychrophiles. Results Core samples obtained from a Cr(VI contaminated aquifer at the Hanford facility in Washington were enriched in Vogel Bonner medium at 10 Centigrade with 0, 25, 50, 100, 200, 400 and 1000 mg/l Cr(VI. The extent of Cr(VI reduction was evaluated using the diphenyl carbazide assay. Resistance to Cr(VI up to and including 1000 mg/l Cr(VI was observed in the consortium experiments. Reduction was slow or not observed at and above 100 mg/l Cr(VI using the enrichment consortium. Average time to complete reduction of Cr(VI in the 30 and 60 mg/l Cr(VI cultures of the consortium was 8 and 17 days, respectively at 10 Centigrade. Lyophilized consortium cells did not demonstrate adsorption of Cr(VI over a 24 hour period. Successful isolation of a Cr(VI reducing organism (designated P4 from the consortium was confirmed by 16S rDNA amplification and sequencing. Average time to complete reduction of Cr(VI at 10 Centigrade in the 25 and 50 mg/l Cr(VI cultures of the isolate P4 was 3 and 5 days, respectively. The 16S rDNA sequence from isolate P4 identified this organism as a strain of Arthrobacter aurescens, a species that has not previously been shown to be capable of low temperature Cr(VI reduction. Conclusion A. aurescens, indigenous to the subsurface, has the potential to be a predominant metal reducer in enhanced, in situ subsurface bioremediation efforts involving Cr(VI and possibly other heavy metals and radionuclides.

  12. PLASMID-ENCODED PHTHALATE CATABOLIC PATHWAY IN ARTHROBACTER KEYSERI 12B: BIOTRANSFORMATIONS OF 2-SUBSTITUTED BENZOATES AND THEIR USE IN CLONING AND CHARACTERIZATION OF PHTHALATE CATABOLISM GENES AND GENE PRODUCTS

    Science.gov (United States)

    Several 2-substituted benzoates (including 2-trifluoromethyl-, 2-chloro-, 2-bromo-, 2-iodo-, 2-nitro-, 2-methoxy-, and 2-acetyl-benzoates) were converted by phthalate-grown Arthrobacter keyseri 12B to the corresponding 2-substituted 3,4-dihydroxybenzoates (protocatechuates)...

  13. Improvement of endophytic Azospirillum colonization by co-inoculation with Cellulomonas Uda ATCC 491

    Directory of Open Access Journals (Sweden)

    Mohammad Javad Mehdipour Moghaddam

    2014-04-01

    Full Text Available Introduction: Most of the plant growth promoting rhizobacteria (PGPR such as Azopirillum if accompanied with strong cellulase producing bacteria such as Cellulomonas, their colonization may be increased and their host plants growth improved. Materials and methods: Six endophytic Azospirilla which isolated from three rice and three wheat cultivars and also one strain from commercial biofertilizer (Green Biotech Co., identified by biochemical tests and 16S rDNA analysis and were studied on the basis of cellulase, pectinase and auxin production and also their chemotaxis toward rice and wheat cultivars exudates was investigated. Two cellulase positive (A5 and A6 and two negative (A2 and A3 strains were selected and their interaction with C. uda ATCC 491 on auxin production and colonization on roots were compared. Results: This study showed that none of the strains had pectinase activity, but the strain isolated from rice had more Carboxy methyl cellulase (CMCase activity. Selected isolates and C. uda ATCC 491 showed chemotaxis toward roots exudates. In most of the isolates, rate of auxin production increased by coculture with C. uda ATCC 491. Also, it was determined that C. uda ATCC 491 promoted the colonization of Azospirillum without or with cellulase activity on rice and wheat roots, respectively. Discussion and conclusion: Co-inoculation Azospirillum with C. uda ATCC 491 improves plant root system due to stimulation or additive effect of auxin production and cellulase activity, followed by more uptakes of water and minerals by roots. Also, it raises the number of colonization niches for useful bacteria such as Azospirillum and finally quantitative and qualitative plant parameters.

  14. Characterization of the binding of Actinomyces naeslundii (ATCC 12104) and Actinomyces viscosus (ATCC 19246) to glycosphingolipids, using a solid-phase overlay approach

    Energy Technology Data Exchange (ETDEWEB)

    Stroemberg, N.K.; Karlsson, K.A. (Univ. of Goeteborg (Sweden))

    1990-07-05

    Actinomyces naeslundii (ATCC 12104) and Actinomyces viscosus (ATCC 19246) were radiolabeled externally (125I) or metabolically (35S) and analyzed for their ability to bind glycosphingolipids separated on thin layer chromatograms or coated in microtiter wells. Two binding properties were found and characterized in detail. (i) Both bacteria showed binding to lactosylceramide (LacCer) in a fashion similar to bacteria characterized earlier. The activity of free LacCer was dependent on the ceramide structure; species with 2-hydroxy fatty acid and/or a trihydroxy base were positive, while species with nonhydroxy fatty acid and a dihydroxy base were negative binders. Several glycolipids with internal lactose were active but only gangliotriaosylceramide and gangliotetraosylceramide were as active as free LacCer. The binding to these three species was half-maximal at about 200 ng of glycolipid and was not blocked by preincubation of bacteria with free lactose or lactose-bovine serum albumin. (ii) A. naeslundii, unlike A. viscosus, showed a superimposed binding concluded to be to terminal or internal GalNAc beta and equivalent to a lactose-inhibitable specificity previously analyzed by other workers. Terminal Gal beta was not recognized in several glycolipids, although free Gal and lactose were active as soluble inhibitors. The binding was half-maximal at about 10 ng of glycolipid. A glycolipid mixture prepared from a scraping of human buccal epithelium contained an active glycolipid with sites for both binding specificities.

  15. Bio desulfurization of a system containing synthetic fuel by rhodococcus erythropolis ATCC 4277; Remocao de compostos sulfurosos de sitema bifasico contendo combustivel sintetico por Rhodococcus erythropolis ATCC 4277

    Energy Technology Data Exchange (ETDEWEB)

    Maass, Danielle; Souza, Antonio Augusto Ulson de; Souza, Selene Maria de Arruda Guelli Ulson de [Universidade Federal de Santa Catarina (UFSC), SC (Brazil)

    2012-07-01

    For decades the burning of fossil fuels released a lot of pollutants in the atmosphere. Among the most harmful is sulfur dioxide (SO{sub 2}), which reacts with the moisture in the air and turns into sulfuric acid, being the main cause of acid rain. Acid rain is very harmful to animal and plant kingdoms; accelerates the corrosion's processes of buildings and monuments, and causes serious health problems for humans. As a result, many countries have reformed their legislation to require the sale of fuels with very low sulfur content. The existing processes of desulfurization are not capable of removing sulfur so low. Therefore, there has developed a new process called bio desulfurization. In this process, the degradation of sulfur occurs through the action of microorganisms that act as catalysts. The bacterium Rhodococcus erythropolis has emerged as one of the most promising for bio desulfurization because it removes the sulfur without breaking the benzene rings, thereby maintaining the potential energy of the same. Using dibenzothiophene as a model of sulfur compounds, the products of the bio desulfurization process are 2- hydroxybiphenyl and sulfate. In this study we sought to examine the desulfurizing capacity of national Rhodococcus erythropolis strain ATCC4277 in a batch reactor using concentrations of organic phase (n-dodecane) of 20 and 80% (v/v). Rhodococcus erythropolis ATCC4277 was capable of degrading DBT in 93.3 and 98.0% in the presence of 20 and 80% (v/v) of synthetic fuel, respectively. (author)

  16. Production of β -cyclodextrin from pH and thermo stable Cyclodextrin Glycosyl Transferase, obtained from Arthrobacter mysorens and its evaluation as a drug carrier for Irbesartan.

    Science.gov (United States)

    Rajesh, Y; Narayanan, K; Reddy, M Sreenivasa; Bhaskar, Vijaya K; Shenoy, G Gautham; Subrahmanyam, V M; Rao, J Venkata

    2015-01-01

    Cyclodextrins (CDs) are carrier molecules produced by cyclization of α-1,4-glucans by Cyclodextrin Glycosyl Transferase (CGTase). These torus shaped molecules have hydrophobic cavity and hydrophilic shell making them useful in pharmaceutical, food, textile, pesticide and cosmetic industries. In this study, culture conditions for the production of CGTase by organism belonging to Arthrobacter genus obtained from a paddy field soil were optimized by single parameter mode. Soluble starch, yeast extract and magnesium sulphate played an important role in CGTase production. Percentage increase in CGTase yield under optimized conditions was 396.77%. The enzyme precipitated by 60% ammonium sulphate was purified using DEAE-sepharose. The molecular weight of the purified protein as determined by SDS-PAGE was 75 kDa. Purified CGTase was thermostable and stable over a wide pH range. Dissolution studies on β -cyclodextrin-Irbesartan complex revealed that β -CDs formed were useful in preparing immediate release oral dosage forms. PMID:25901452

  17. A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties

    Directory of Open Access Journals (Sweden)

    Kur Józef

    2009-07-01

    Full Text Available Abstract Background The development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production. Results In this article, we present a new β-D-galactosidase as a candidate to be applied in the above mentioned biotechnological processes. The gene encoding this β-D-galactosidase has been isolated from the genomic DNA library of Antarctic bacterium Arthrobacter sp. 32c, sequenced, cloned, expressed in Escherichia coli and Pichia pastoris, purified and characterized. 27 mg of β-D-galactosidase was purified from 1 L of culture with the use of an intracellular E. coli expression system. The protein was also produced extracellularly by P. pastoris in high amounts giving approximately 137 mg and 97 mg of purified enzyme from 1 L of P. pastoris culture for the AOX1 and a constitutive system, respectively. The enzyme was purified to electrophoretic homogeneity by using either one step- or a fast two step- procedure including protein precipitation and affinity chromatography. The enzyme was found to be active as a homotrimeric protein consisting of 695 amino acid residues in each monomer. Although, the maximum activity of the enzyme was determined at pH 6.5 and 50°C, 60% of the maximum activity of the enzyme was determined at 25°C and 15% of the maximum activity was detected at 0°C. Conclusion The properties of Arthrobacter sp. 32cβ-D-galactosidase suggest that this enzyme could be useful for low-cost, industrial conversion of lactose into galactose and glucose in milk products and could be an interesting alternative for the production of ethanol from lactose-based feedstock.

  18. Phosphoketolase Pathway Dominates in Lactobacillus reuteri ATCC 55730 Containing Dual Pathways for Glycolysis▿

    OpenAIRE

    Årsköld, Emma; Lohmeier-Vogel, Elke; Cao, Rong; Roos, Stefan; Rådström, Peter; van Niel, Ed W. J.

    2007-01-01

    Metabolic flux analysis indicated that the heterofermentative Lactobacillus reuteri strain ATCC 55730 uses both the Embden-Meyerhof pathway (EMP) and phosphoketolase pathway (PKP) when glucose or sucrose is converted into the three-carbon intermediate stage of glycolysis. In all cases studied, the main flux is through the PKP, while the EMP is used as a shunt. In the exponential growth phase, 70%, 73%, and 84% of the flux goes through the PKP in cells metabolizing (i) glucose plus fructose, (...

  19. Phosphoketolase pathway dominates in Lactobacillus reuteri ATCC 55730 containing dual pathways for glycolysis

    OpenAIRE

    Årsköld, Emma; Lohmeier-Vogel, Elke; Cao, Rong; Roos, Stefan; Rådström, Peter; van Niel, Ed

    2008-01-01

    Metabolic flux analysis indicated that the heterofermentative Lactobacillus reuteri strain ATCC 55730 uses both the Embden-Meyerhof pathway (EMP) and phosphoketolase pathway (PKP) when glucose or sucrose is converted into the three-carbon intermediate stage of glycolysis. In all cases studied, the main flux is through the PKP, while the EMP is used as a shunt. In the exponential growth phase, 70%, 73%, and 84% of the flux goes through the PKP in cells metabolizing (i) glucose plus fructose, (...

  20. Genome sequence of the methanotrophic Alphaproteobacterium, Methylocystis sp. Rockwell (ATCC 49242)

    Energy Technology Data Exchange (ETDEWEB)

    Stein, Lisa Y. [University of Alberta, Edmondton, Canada; Bringel, Francoise O. [University of Strasbourg; DiSpiritto, Alan A. [University of Iowa; Han, Sukkyun [University of Alberta, Edmondton, Canada; Jetten, MSM [Radboud University Nijmegen, The Netherlands; Kalyuzhnaya, Marina G. [University of Washington, Seattle; Kits, K. Dimitri [University of Alberta, Edmondton, Canada; Klotz, Martin G [University of Louisville, Louisville; Op den Camp, HJM [Radboud University Nijmegen, The Netherlands; Semrau, Jeremy D. [University of Michigan; Vuilleumier, Stephane [University of Strasbourg; Bruce, David [Los Alamos National Laboratory (LANL); Cheng, Jan-Fang [U.S. Department of Energy, Joint Genome Institute; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Davenport, Karen W. [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Hauser, Loren John [ORNL; Lajus, Aurelie [Genoscope/Centre National de la Recherche Scientifique-Unite Mixte de Recherche; Land, Miriam L [ORNL; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Medigue, Claudine [Genoscope/Centre National de la Recherche Scientifique-Unite Mixte de Recherche; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute

    2011-01-01

    Methylocystis sp. strain Rockwell (ATCC 49242) is an aerobic methane-oxidizing Alphaproteobacterium isolated from an aquifer in southern California. Unlike most methanotrophs in the Methylocystaceae family, this strain has a single pmo operon encoding particulate methane monooxygenase and no evidence of the genes encoding soluble methane monooxygenase. This is the first reported genome sequence of a member of the Methylocystis species of the Methylocystaceae family in the order Rhizobiales.

  1. Sorbitol Production By Zymomonas Mobilis ATCC 29191 In Medium Of Sucrose Pre-Treated With Invertase

    Directory of Open Access Journals (Sweden)

    Maria Antonia Pedrine Colabone Celligoi

    2002-01-01

    Full Text Available Sorbitol production by Zymomonas mobilis ATCC 29191 in medium of sucrose pre-treated with invertase was studied. The best results were obtained when the medium was pre-treated with invertase as sorbitol production of 41,39 g/L and a productivity of 1,72 g/L.h-1 in 24 hours of fermentation. The invertase addition in the fermentation broth increased 72,17% in the sorbitol formation.

  2. Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

    OpenAIRE

    Gallo, Giuseppe; Renzone, Giovanni; Palazzotto, Emilia; Monciardini, Paolo; Arena, Simona; Faddetta, Teresa; Giardina, Anna; Alduina, Rosa; Weber, Tilmann; Sangiorgi, Fabio; Russo, Alessandro; Spinelli, Giovanni; Sosio, Margherita; Scaloni, Andrea; Puglia, Anna Maria

    2016-01-01

    Background The filamentous actinomycete Microbispora ATCC-PTA-5024 produces the lantibiotic NAI-107, which is an antibiotic peptide effective against multidrug-resistant Gram-positive bacteria. In actinomycetes, antibiotic production is often associated with a physiological differentiation program controlled by a complex regulatory and metabolic network that may be elucidated by the integration of genomic, proteomic and bioinformatic tools. Accordingly, an extensive evaluation of the proteomi...

  3. Agroindustrial Byproducts For The Production Of Hyaluronic Acid By Streptococcus Zooepidemicus ATCC 39920

    OpenAIRE

    Nicole Caldas Pan; Josiane Alessandra Vignoli; Cristiani Baldo; Hanny Cristina Braga Pereira; Rui Srgio dos Santos Ferreira da Silva; Maria Antonia Pedrine Colabone Celligoi

    2015-01-01

    Abstract Agroindustrial derivatives are alternative nutritional sources employed in bioprocesses that reduce costs and corroborate with social sustainability. In this study alternative carbon sugarcane juice sugarcane molasses and soy molasses and nitrogen sources corn steep liquor soy protein and whey protein were evaluated for hyaluronic acid production by Streptococcus zooepidemicus ATCC 39920. The medium containing sugarcane molasses archived high yield of hyaluronic acid 0.066 g.g-1 when...

  4. Sorbitol Production By Zymomonas Mobilis ATCC 29191 In Medium Of Sucrose Pre-Treated With Invertase

    OpenAIRE

    Maria Antonia Pedrine Colabone Celligoi; Viviane Cristina Schiabel; João Batista Buzato; Josiane Alessandra Vignoli; Márcio de Barros

    2002-01-01

    Sorbitol production by Zymomonas mobilis ATCC 29191 in medium of sucrose pre-treated with invertase was studied. The best results were obtained when the medium was pre-treated with invertase as sorbitol production of 41,39 g/L and a productivity of 1,72 g/L.h-1 in 24 hours of fermentation. The invertase addition in the fermentation broth increased 72,17% in the sorbitol formation.

  5. Optimization of probiotic lactobacillus casei ATCC 334 production using date powder as carbon source

    OpenAIRE

    Shahravy A.; Tabandeh F.; Bambai B.; Zamanizadeh H.R.; Mizani M.

    2012-01-01

    This study was conducted to optimize culture conditions for economic production of a probiotic bacterium, Lactobacillus casei ATCC 334, in which palm date powder was applied for the first time as a low-cost main carbon source. The effect of eleven factors on bacterial growth was investigated using the Taguchi experimental design, and three factors including palm date powder, tryptone and agitation rate were found to be the most significant parameters. The optimum conditions including da...

  6. Study of nano-fiber cellulose production by Glucanacetobacter xylinum ATCC 10245.

    OpenAIRE

    Norouzian, D.; Farhangi, Ali; Tolooei, S.; Saffari, Z.; Mehrabi, M. R.; Chiani, M.; Ghassemi, S.; Farahnak, M.; Akbarzadeh, Azim

    2011-01-01

    International audience Bacterial Celluloses (BC) are gaining importance in research and commerce due to numerous factors affecting the bacterial cellulose characteristics and application in different industries. The aim of the present study was to produce bacterial cellulose in different media using different cultivation vessels. Bacterial cellulose was produced by static cultivation of Glucanacetobacter xylinum ATCC 10245 in different culture media such as Brain Heart Agar, Luria Bertani ...

  7. Genome sequence of the Bacteroides fragilis phage ATCC 51477-B1

    Directory of Open Access Journals (Sweden)

    Hawkins Shawn A

    2008-08-01

    Full Text Available Abstract The genome of a fecal pollution indicator phage, Bacteroides fragilis ATCC 51477-B1, was sequenced and consisted of 44,929 bases with a G+C content of 38.7%. Forty-six putative open reading frames were identified and genes were organized into functional clusters for host specificity, lysis, replication and regulation, and packaging and structural proteins.

  8. Evaluating Chemical Mitigation of Salmonella Typhimurium ATCC 14028 in Animal Feed Ingredients.

    Science.gov (United States)

    Cochrane, Roger A; Huss, Anne R; Aldrich, Gregory C; Stark, Charles R; Jones, Cassandra K

    2016-04-01

    Salmonella Typhimurium is a potential feed safety hazard in animal feed ingredients. Thermal mitigation of Salmonella spp. during rendering is effective but does not eliminate the potential for cross-contamination. Therefore, the objective of this experiment was to evaluate the effectiveness of chemicals to mitigate postrendering Salmonella Typhimurium ATCC 14028 contamination in rendered proteins over time. Treatments were arranged in a 6 × 4 factorial with six chemical treatments and four rendered protein meals. The chemical treatments included (i) control without chemical treatment, (ii) 0.3% commercial formaldehyde product, (iii) 2% essential oil blend, (iv) 2% medium chain fatty acid blend, (v) 3% organic acid blend, and (vi) 1% sodium bisulfate. The four rendered protein meals included (i) feather meal, (ii) blood meal, (iii) meat and bone meal, and (iv) poultry by-product meal. After matrices were chemically treated, they were inoculated with Salmonella Typhimurium ATCC 14028, stored at room temperature, and enumerated via plate counts on days 0, 1, 3, 7, 14, 21, and 42 postinoculation. The Salmonella concentration in ingredients treated with medium chain fatty acid and commercial formaldehyde were similar to one another (P = 0.23) but were 2 log lower than the control (P organic acids and essential oils also had lower Salmonella concentrations than the control (P poultry by-product meal (P acids or a commercial formaldehyde product were most effective at mitigating Salmonella Typhimurium ATCC 14028 in rendered protein meals. PMID:27052874

  9. Dynamic proteome analysis of Cyanothece sp. ATCC 51142 under constant light

    Energy Technology Data Exchange (ETDEWEB)

    Aryal, Uma K.; Stockel, Jana; Welsh, Eric A.; Gritsenko, Marina A.; Nicora, Carrie D.; Koppenaal, David W.; Smith, Richard D.; Pakrasi, Himadri B.; Jacobs, Jon M.

    2012-02-03

    Understanding the dynamic nature of protein abundances provides insights into protein turnover not readily apparent from conventional, static mass spectrometry measurements. This level of data is particularly informative when surveying protein abundances in biological systems subjected to large perturbations or alterations in environment such as cyanobacteria. Our current analysis expands upon conventional proteomic approaches in cyanobacteria by measuring dynamic changes of the proteome using a 13C15N-L-leucine metabolic labeling in Cyanothece ATCC51142. Metabolically labeled Cyanothece ATCC51142 cells grown under nitrogen sufficient conditions in continuous light were monitored longitudinally for isotope incorporation over a 48 h period, revealing 422 proteins with dynamic changes in abundances. In particular, proteins involved in carbon fixation, pentose phosphate pathway, cellular protection, redox regulation, protein folding, assembly and degradation showed higher levels of isotope incorporation suggesting that these biochemical pathways are important for growth under non-diazotrophic conditions. Calculation of relative isotope abundances (RIA) values allowed to measure actual active protein synthesis over time for different biochemical pathways under non-diazotrophic conditions. Overall results demonstrated the utility of 'non-steady state' pulsed metabolic labeling for systems-wide dynamic quantification of the proteome in Cyanothece ATCC51142 that can also be applied to other cyanobacteria.

  10. Genome-scale reconstruction of metabolic networks of Lactobacillus casei ATCC 334 and 12A.

    Directory of Open Access Journals (Sweden)

    Elena Vinay-Lara

    Full Text Available Lactobacillus casei strains are widely used in industry and the utility of this organism in these industrial applications is strain dependent. Hence, tools capable of predicting strain specific phenotypes would have utility in the selection of strains for specific industrial processes. Genome-scale metabolic models can be utilized to better understand genotype-phenotype relationships and to compare different organisms. To assist in the selection and development of strains with enhanced industrial utility, genome-scale models for L. casei ATCC 334, a well characterized strain, and strain 12A, a corn silage isolate, were constructed. Draft models were generated from RAST genome annotations using the Model SEED database and refined by evaluating ATP generating cycles, mass-and-charge-balances of reactions, and growth phenotypes. After the validation process was finished, we compared the metabolic networks of these two strains to identify metabolic, genetic and ortholog differences that may lead to different phenotypic behaviors. We conclude that the metabolic capabilities of the two networks are highly similar. The L. casei ATCC 334 model accounts for 1,040 reactions, 959 metabolites and 548 genes, while the L. casei 12A model accounts for 1,076 reactions, 979 metabolites and 640 genes. The developed L. casei ATCC 334 and 12A metabolic models will enable better understanding of the physiology of these organisms and be valuable tools in the development and selection of strains with enhanced utility in a variety of industrial applications.

  11. Cloning, Purification and Characterization of the Collagenase ColA Expressed by Bacillus cereus ATCC 14579.

    Science.gov (United States)

    Abfalter, Carmen M; Schönauer, Esther; Ponnuraj, Karthe; Huemer, Markus; Gadermaier, Gabriele; Regl, Christof; Briza, Peter; Ferreira, Fatima; Huber, Christian G; Brandstetter, Hans; Posselt, Gernot; Wessler, Silja

    2016-01-01

    Bacterial collagenases differ considerably in their structure and functions. The collagenases ColH and ColG from Clostridium histolyticum and ColA expressed by Clostridium perfringens are well-characterized collagenases that cleave triple-helical collagen, which were therefore termed as ´true´ collagenases. ColA from Bacillus cereus (B. cereus) has been added to the collection of true collagenases. However, the molecular characteristics of B. cereus ColA are less understood. In this study, we identified ColA as a secreted true collagenase from B. cereus ATCC 14579, which is transcriptionally controlled by the regulon phospholipase C regulator (PlcR). B. cereus ATCC 14579 ColA was cloned to express recombinant wildtype ColA (ColAwt) and mutated to a proteolytically inactive (ColAE501A) version. Recombinant ColAwt was tested for gelatinolytic and collagenolytic activities and ColAE501A was used for the production of a polyclonal anti-ColA antibody. Comparison of ColAwt activity with homologous proteases in additional strains of B. cereus sensu lato (B. cereus s.l.) and related clostridial collagenases revealed that B. cereus ATCC 14579 ColA is a highly active peptidolytic and collagenolytic protease. These findings could lead to a deeper insight into the function and mechanism of bacterial collagenases which are used in medical and biotechnological applications. PMID:27588686

  12. Genome sequence and plasmid transformation of the model high-yield bacterial cellulose producer Gluconacetobacter hansenii ATCC 53582

    OpenAIRE

    Michael Florea; Benjamin Reeve; James Abbott; Freemont, Paul S.; Tom Ellis

    2016-01-01

    Bacterial cellulose is a strong, highly pure form of cellulose that is used in a range of applications in industry, consumer goods and medicine. Gluconacetobacter hansenii ATCC 53582 is one of the highest reported bacterial cellulose producing strains and has been used as a model organism in numerous studies of bacterial cellulose production and studies aiming to increased cellulose productivity. Here we present a high-quality draft genome sequence for G. hansenii ATCC 53582 and find that in ...

  13. Effects of Cd (Ⅱ) on the physico-biochemical behaviors of Arthrobacter sp.and Bacillus sp.%镉胁迫对节杆菌(Arthrobacter sp.)和芽孢杆菌(Bacillus sp.)生理生化特性的影响

    Institute of Scientific and Technical Information of China (English)

    杜瑞英

    2012-01-01

    将节杆菌和芽孢杆菌分别暴露于不同质量比的镉溶液中,进行不同暴露时间的急性毒性试验.结果表明,20 mg/kg的Cd(Ⅱ)处理会使节杆菌中还原型谷胱甘肽(GSH)质量浓度显著降低,细胞膜脂质过氧化产物——硫代巴比妥酸活性物质(TBARS)浓度显著升高;0.2mg/kg的Cd(Ⅱ)处理会使芽孢杆菌中GSH质量浓度、过氧化氢酶(CAT)活性显著降低.节杆菌的可溶性蛋白、可溶性糖质量浓度随暴露时间延长而减少,GSH质量浓度、CAT和超氧化物歧化酶(SOD)活性随暴露时间延长而增加,TBARS浓度呈先减后增的变化趋势;芽孢杆菌的可溶性蛋白质量浓度、CAT活性和TBARS浓度呈先增后减的变化趋势,可溶性糖、GSH质量浓度和SOD活性呈先减后增的变化趋势.镉处理对节杆菌和芽孢杆菌具有一定的胁迫作用,两种菌通过启动不同的抗性系统来抵抗外界胁迫.%This paper is engaged in the study of the effects of Cd(II) on the physico-biochemical behaviors of Arthrobacter sp. through tox-icity testing. The toxicity test has been arranged by putting the an-tioxidant system of Arthrobacter sp. and Bacillus sp. under the exposure to different concentrations of Cd(II) for different lengths of testing time. The results of our testing show that Cd(II) has a strong impact on the antioxidant system of Arthrobacter sp. and Bacillus sp. . However, it is possible to reduce GSH content and increase TBARS content of Arthrobacter sp. significantly by letting Cd(II) treated with 20 mg/kg. Our testing proves that it is possible to reduce the GSH content and CAT activity of Bacillus sp. significantly by treating Cd(II) with 0.2 mg/kg, for the Arthrobacter sp. , the soluble protein and soluble sugar content can be reduced through prolonging the time of exposure. On the contrary, it is also possible to increase the GSH content and the CAT & SOD activity by means of prolonging the exposure, with the TBARS content

  14. Efecto del Cromo Hexavalente y Trivalente sobre el Crecimiento de Escherichia coli ATCC 35218 Effects of Hexavalent and Trivalent Chromium on the Growth of Escherichia coli ATCC 35218

    Directory of Open Access Journals (Sweden)

    Ricardo R Azario

    2010-01-01

    Full Text Available Se estudió el efecto de cromo (VI y (III sobre el crecimiento de Escherichia coli ATCC 35218 con el fin de determinar la toxicidad de estas especies químicas. El estudio fue realizado en dos condiciones experimentales: soluciones unimetal de cromo (III o VI y en soluciones multimetal conteniendo además plomo y cadmio. Se observa que el cromo hexavalente inhibe el crecimiento de Escherichia coli, cuando se encuentra en un rango de concentración de 25 a 100 ppm, similar al de efluentes industriales, y que dicho efecto es potenciado por la presencia de plomo, que per se no modifica la viabilidad bacteriana. Por otro lado, las concentraciones bajas de cromo (VI, 0.05 - 5 ppm no alteran el crecimiento pero producen una estimulación en presencia de plomo o cadmio. La forma trivalente de cromo no modifica el crecimiento bacteriano a concentraciones bajas (25 a 100 ppm pero causa una estimulación a concentraciones más altas (200 a 400 ppm.The effect of chromium (III and (VI on the growth of Escherichia coli ATCC 35218 was studied to determine the toxicity of these chemical species. The study was performed in two experimental conditions: single chromium solutions (III or VI and multimetal solutions containing chromium and either lead or cadmium. Hexavalent chromium, at concentrations from 25 to 100 ppm, similar to those found in industrial effluents, inhibits the growth of Escherichia coli. This inhibitory effect is increased by the presence of lead, which does not modify per se the bacterial viability. On the other hand, low concentrations of chromium (VI, 0.05-5 ppm do not alter bacterial growth but cause stimulation in the presence of either lead or cadmium. The trivalent form of chromium does not modify the bacterial growth at low concentrations (25 to 100 ppm but causes stimulation at high concentrations (200 to 400 ppm.

  15. Investigation of the Amycolatopsis sp. strain ATCC 39116 vanillin dehydrogenase and its impact on the biotechnical production of vanillin.

    Science.gov (United States)

    Fleige, Christian; Hansen, Gunda; Kroll, Jens; Steinbüchel, Alexander

    2013-01-01

    The actinomycete Amycolatopsis sp. strain ATCC 39116 is capable of synthesizing large amounts of vanillin from ferulic acid, which is a natural cell wall component of higher plants. The desired intermediate vanillin is subject to undesired catabolism caused by the metabolic activity of a hitherto unknown vanillin dehydrogenase (VDH(ATCC 39116)). In order to prevent the oxidation of vanillin to vanillic acid and thereby to obtain higher yields and concentrations of vanillin, the responsible vanillin dehydrogenase in Amycolatopsis sp. ATCC 39116 was investigated for the first time by using data from our genome sequence analysis and further bioinformatic approaches. The vdh gene was heterologously expressed in Escherichia coli, and the encoded vanillin dehydrogenase was characterized in detail. VDH(ATCC 39116) was purified to apparent electrophoretic homogeneity and exhibited NAD(+)-dependent activity toward vanillin, coniferylaldehyde, cinnamaldehyde, and benzaldehyde. The enzyme showed its highest level of activity toward vanillin at pH 8.0 and at a temperature of 44°C. In a next step, a precise vdh deletion mutant of Amycolatopsis sp. ATCC 39116 was generated. The mutant lost its ability to grow on vanillin and did not show vanillin dehydrogenase activity. A 2.3-times-higher vanillin concentration and a substantially reduced amount of vanillic acid occurred with the Amycolatopsis sp. ATCC 39116 Δvdh::Km(r) mutant when ferulic acid was provided for biotransformation in a cultivation experiment on a 2-liter-bioreactor scale. Based on these results and taking further metabolic engineering into account, the Amycolatopsis sp. ATCC 39116 Δvdh::Km(r) mutant represents an optimized and industrially applicable platform for the biotechnological production of natural vanillin.

  16. Regio-specific Microbial Hydroxylation of Phytolaccagenin by Streptomyces griseus ATCC 13273

    Institute of Scientific and Technical Information of China (English)

    QIAN, Liwu; ZHANG, Jian; LIU, Jihua; YU, Boyang

    2009-01-01

    Microbial transformation of one oleane-type pentacyclic triterpene aglycone, phytolaccagenin (2β,3β,23-trihy- droxy-olean-12-ene-28,30-dioic acid 30-methyl ester) by Streptomyces griseus ATCC 13273 was investigated for developing new bioactive derivatives. A new oxidized metabolite, through the regio-specific hydroxylation on the C-29 methyl group, was obtained from the preparative-scale biotransformation with a standard two-stage fermenta- tion protocol. The metabolite was identified as 2β,3β,23,29-tetrahydroxy-olean-12-ene-28,30-dioic acid 30-methyl ester by mass and 2D-NMR spectra.

  17. DNA Sequence and Comparison of Virulence Plasmids from Rhodococcus equi ATCC 33701 and 103

    OpenAIRE

    Takai, Shinji; Hines, Stephen A.; Sekizaki, Tsutomu; Nicholson, Vivian M.; Alperin, Debra A.; Osaki, Makoto; Takamatsu, Daisuke; Nakamura, Mutsu; SUZUKI, KAYO; Ogino, Nobuko; Kakuda, Tsutomu; Dan, Hanhong; Prescott, John F.

    2000-01-01

    The virulence plasmids of the equine virulent strains Rhodococcus equi ATCC 33701 and 103 were sequenced, and their genetic structure was analyzed. p33701 was 80,610 bp in length, and p103 was 1 bp shorter; their sequences were virtually identical. The plasmids contained 64 open reading frames (ORFs), 22 of which were homologous with genes of known function and 3 of which were homologous with putative genes of unknown function in other species. Putative functions were assigned to five ORFs ba...

  18. Specificity of an extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine beta-casein.

    OpenAIRE

    Rattray, F P; Fox, P. F.; Healy, A.

    1997-01-01

    The specificity of the extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine beta-casein was studied. Hydrolysis was monitored over time by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and urea-PAGE. The major pH 4.6-soluble peptides were isolated by high-performance liquid chromatography and identified by N-terminal amino acid sequencing and mass spectrometry. The major sites of hydrolysis were Ser-18-Ser-19, Glu-20-Glu-21, Gln-56-Ser-57, Gln-72-Asn-73, ...

  19. Purification and Characterization of an Extracellular Proteinase from Brevibacterium linens ATCC 9174

    OpenAIRE

    Rattray, F P; Bockelmann, W; Fox, P. F.

    1995-01-01

    An extracellular serine proteinase from Brevibacterium linens ATCC 9174 was purified to homogeneity. pH and temperature optima were 8.5 and 50(deg)C, respectively. The results for the molecular mass of the proteinase were 56 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 126 kDa by gel filtration, indicating that the native enzyme exists as a dimer. Mg(sup2+) and Ca(sup2+) activated the proteinase, as did NaCl; however, Hg(sup2+), Fe(sup2+), and Zn(sup2+) caused strong i...

  20. Specificity of an extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine alpha s1-casein.

    OpenAIRE

    Rattray, F P; Fox, P. F.; Healy, A.

    1996-01-01

    The specificity of the extracellular proteinase from Brevibacterium linens ATCC 9174 on bovine alpha s1-casein was studied. Hydrolysis was monitored over time by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and urea-PAGE. The major pH 4.6-soluble peptides were isolated by high-performance liquid chromatography and identified by N-terminal amino acid sequencing and mass spectrometry. The time course of peptide formation indicated that His-8-Gln-9, Ser-161-Gly-162, and eithe...

  1. SpoIIE Regulates Sporulation but Does Not Directly Affect Solventogenesis in Clostridium acetobutylicum ATCC 824

    OpenAIRE

    Scotcher, Miles C.; Bennett, George N.

    2005-01-01

    Using gene expression reporter vectors, we examined the activity of the spoIIE promoter in wild-type and spo0A-deleted strains of Clostridium acetobutylicum ATCC 824. In wild-type cells, the spoIIE promoter is active in a transient manner during late solventogenesis, but in strain SKO1, where the sporulation initiator spo0A is disrupted, no spoIIE promoter activity is detectable at any stage of growth. Strains 824(pMSpo) and 824(pASspo) were created to overexpress spoIIE and to decrease spoII...

  2. Reducing the Bitterness of Tuna (Euthynnus pelamis) Dark Meat with Lactobacillus casei subsp. casei ATCC 393

    OpenAIRE

    Bertoldi, Fabiano Cleber; Ernani S. Sant’Anna; Luiz H. Beirão

    2004-01-01

    During the process of canning tuna fish, considerable amounts of dark tuna meat are left over because of its bitterness, which are then used in the production of animal food. Fermentation with Lactobacillus casei subsp. casei ATCC 393 was used as an alternative to reduce this bitter taste. Samples of meat were prepared, vacuum packed and then stored at –18 °C. The frozen dark meat was used immediately after defrosting and the experiment was carried out with 2 and 4 % of NaCl with the addition...

  3. Evaluating Chemical Mitigation of Salmonella Typhimurium ATCC 14028 in Animal Feed Ingredients.

    Science.gov (United States)

    Cochrane, Roger A; Huss, Anne R; Aldrich, Gregory C; Stark, Charles R; Jones, Cassandra K

    2016-04-01

    Salmonella Typhimurium is a potential feed safety hazard in animal feed ingredients. Thermal mitigation of Salmonella spp. during rendering is effective but does not eliminate the potential for cross-contamination. Therefore, the objective of this experiment was to evaluate the effectiveness of chemicals to mitigate postrendering Salmonella Typhimurium ATCC 14028 contamination in rendered proteins over time. Treatments were arranged in a 6 × 4 factorial with six chemical treatments and four rendered protein meals. The chemical treatments included (i) control without chemical treatment, (ii) 0.3% commercial formaldehyde product, (iii) 2% essential oil blend, (iv) 2% medium chain fatty acid blend, (v) 3% organic acid blend, and (vi) 1% sodium bisulfate. The four rendered protein meals included (i) feather meal, (ii) blood meal, (iii) meat and bone meal, and (iv) poultry by-product meal. After matrices were chemically treated, they were inoculated with Salmonella Typhimurium ATCC 14028, stored at room temperature, and enumerated via plate counts on days 0, 1, 3, 7, 14, 21, and 42 postinoculation. The Salmonella concentration in ingredients treated with medium chain fatty acid and commercial formaldehyde were similar to one another (P = 0.23) but were 2 log lower than the control (P < 0.05). Ingredients treated with organic acids and essential oils also had lower Salmonella concentrations than the control (P < 0.05). Time also played a significant role in Salmonella mitigation, because all days except days 14 and 21 (P = 0.92) differed from one another. Rendered protein matrix also affected Salmonella stability, because concentrations in meat and bone meal and blood meal were similar to one another (P = 0.36) but were greater than levels in feather meal and poultry by-product meal (P < 0.05). In summary, chemical treatment and time both mitigated Salmonella Typhimurium ATCC 14028, but their effectiveness was matrix dependent. Time and chemical treatment with medium

  4. Purification and Characterization of an Extracellular Proteinase from Brevibacterium-Linens ATCC-9174

    DEFF Research Database (Denmark)

    Rattray, F P; Bockelmann, W; Fox, P F

    1995-01-01

    An extracellular serine proteinase from Brevibacterium linens ATCC 9174 was purified to homogeneity. pH and temperature optima were 8,5 and 50 degrees C, respectively. The results for the molecular mass of the proteinase were 56 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis...... and 126 kDa by gel filtration, indicating that the native enzyme exists as a dimer. Mg2+ and Ca2+ activated the proteinase, as did NaCl; however, Hg2+ Fe2+, and Zn2+ caused strong inhibition. The sequence of the first 20 N-terminal amino acids was NH2-Ala-Lys- Asn...

  5. Multicenter Investigation of Gepotidacin (GSK2140944) Agar Dilution Quality Control Determinations for Neisseria gonorrhoeae ATCC 49226.

    Science.gov (United States)

    Jones, Ronald N; Fedler, Kelley A; Scangarella-Oman, Nicole E; Ross, James E; Flamm, Robert K

    2016-07-01

    Gepotidacin, a novel triazaacenaphthylene antibacterial agent, is the first in a new class of type IIA topoisomerase inhibitors with activity against many biothreat and conventional pathogens, including Neisseria gonorrhoeae To assist ongoing clinical studies of gepotidacin to treat gonorrhea, a multilaboratory quality assurance investigation determined the reference organism (N. gonorrhoeae ATCC 49226) quality control MIC range to be 0.25 to 1 μg/ml (88.8% of gepotidacin MIC results at the 0.5 μg/ml mode). PMID:27161642

  6. Assessment of CcpA-mediated catabolite control of gene expression in Bacillus cereus ATCC 14579

    Directory of Open Access Journals (Sweden)

    Buist Girbe

    2008-04-01

    Full Text Available Abstract Background The catabolite control protein CcpA is a transcriptional regulator conserved in many Gram-positives, controlling the efficiency of glucose metabolism. Here we studied the role of Bacillus cereus ATCC 14579 CcpA in regulation of metabolic pathways and expression of enterotoxin genes by comparative transcriptome analysis of the wild-type and a ccpA-deletion strain. Results Comparative analysis revealed the growth performance and glucose consumption rates to be lower in the B. cereus ATCC 14579 ccpA deletion strain than in the wild-type. In exponentially grown cells, the expression of glycolytic genes, including a non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase that mediates conversion of D-glyceraldehyde 3-phosphate to 3-phospho-D-glycerate in one single step, was down-regulated and expression of gluconeogenic genes and genes encoding the citric acid cycle was up-regulated in the B. cereus ccpA deletion strain. Furthermore, putative CRE-sites, that act as binding sites for CcpA, were identified to be present for these genes. These results indicate CcpA to be involved in the regulation of glucose metabolism, thereby optimizing the efficiency of glucose catabolism. Other genes of which the expression was affected by ccpA deletion and for which putative CRE-sites could be identified, included genes with an annotated function in the catabolism of ribose, histidine and possibly fucose/arabinose and aspartate. Notably, expression of the operons encoding non-hemolytic enterotoxin (Nhe and hemolytic enterotoxin (Hbl was affected by ccpA deletion, and putative CRE-sites were identified, which suggests catabolite repression of the enterotoxin operons to be CcpA-dependent. Conclusion The catabolite control protein CcpA in B. cereus ATCC 14579 is involved in optimizing the catabolism of glucose with concomitant repression of gluconeogenesis and alternative metabolic pathways. Furthermore, the results point to metabolic control

  7. INFLUENCE OF HIGH LIGHT INTENSITY ON THE CELLS OF CYANOBACTERIA ANABAENA VARIABILIS SP. ATCC 29413

    Directory of Open Access Journals (Sweden)

    OPRIŞ SANDA

    2012-12-01

    Full Text Available In this article is presented the result of research regardind the effect of high light intensity on the cells of Anabaena variabilis sp. ATCC 29413, the main objective is to study the adaptation of photosynthetic apparatus to light stress. Samples were analyzed in the present of herbicide diuron (DCMU which blocks electron flow from photosystem II and without diuron. During treatment maximum fluorescence and photosystems efficiency are significantly reduced, reaching very low values compared with the blank, as a result of photoinhibition installation. Also by this treatment is shown the importance of the mechanisms by which cells detect the presence of light stress and react accordingly.

  8. Glucose metabolism in the antibiotic producing actinomycete Nonomuraea sp ATCC 39727

    DEFF Research Database (Denmark)

    Gunnarsson, Nina; Bruheim, Per; Nielsen, Jens

    2004-01-01

    The actinomycete Nonomuraea sp. ATCC 39727, producer of the glycopeptide A40926 that is used as precursor for the novel antibiotic dalbavancin, has an unusual carbon metabolism. Glucose is primarily metabolized via the Entner-Doudoroff (ED) pathway, although the energetically more favorable Embden...... - Meyerhof - Parnas (EMP) pathway is present in this organism. Moreover, Nonomuraea utilizes a PPi-dependent phosphofructokinase, an enzyme that has been connected with anaerobic metabolism in eukaryotes and higher plants, but recently has been recognized in several actinomycetes. In order to study its...

  9. Lactobacillus acidophilus ATCC 4356 prevents atherosclerosis via inhibition of intestinal cholesterol absorption in apolipoprotein E-knockout mice.

    Science.gov (United States)

    Huang, Ying; Wang, Jinfeng; Quan, Guihua; Wang, Xiaojun; Yang, Longfei; Zhong, Lili

    2014-12-01

    The objective of this study was to investigate the effect of Lactobacillus acidophilus ATCC 4356 on the development of atherosclerosis in apolipoprotein E-knockout (ApoE(-/-)) mice. Eight-week-old ApoE(-/-) mice were fed a Western diet with or without L. acidophilus ATCC 4356 daily for 16 weeks. L. acidophilus ATCC 4356 protected ApoE(-/-) mice from atherosclerosis by reducing their plasma cholesterol levels from 923 ± 44 to 581 ± 18 mg/dl, likely via a marked decrease in cholesterol absorption caused by modulation of Niemann-Pick C1-like 1 (NPC1L1). In addition, suppression of cholesterol absorption induced reverse cholesterol transport (RCT) in macrophages through the peroxisome proliferator-activated receptor/liver X receptor (PPAR/LXR) pathway. Fecal lactobacillus and bifidobacterium counts were significantly (P small intestine, colon, and feces during the feeding trial. The bacterial levels remained high even after the administration of lactic acid bacteria had been stopped for 2 weeks. These results suggest that administration of L. acidophilus ATCC 4356 can protect against atherosclerosis through the inhibition of intestinal cholesterol absorption. Therefore, L. acidophilus ATCC 4356 may be a potential therapeutic material for preventing the progression of atherosclerosis.

  10. Phosphoketolase pathway dominates in Lactobacillus reuteri ATCC 55730 containing dual pathways for glycolysis.

    Science.gov (United States)

    Arsköld, Emma; Lohmeier-Vogel, Elke; Cao, Rong; Roos, Stefan; Rådström, Peter; van Niel, Ed W J

    2008-01-01

    Metabolic flux analysis indicated that the heterofermentative Lactobacillus reuteri strain ATCC 55730 uses both the Embden-Meyerhof pathway (EMP) and phosphoketolase pathway (PKP) when glucose or sucrose is converted into the three-carbon intermediate stage of glycolysis. In all cases studied, the main flux is through the PKP, while the EMP is used as a shunt. In the exponential growth phase, 70%, 73%, and 84% of the flux goes through the PKP in cells metabolizing (i) glucose plus fructose, (ii) glucose alone, and (iii) sucrose alone, respectively. Analysis of the genome of L. reuteri ATCC 55730 confirmed the presence of the genes for both pathways. Further evidence for the simultaneous operation of two central carbon metabolic pathways was found through the detection of fructose-1,6-bisphosphate aldolase, phosphofructokinase, and phosphoglucoisomerase activities and the presence of phosphorylated EMP and PKP intermediates using in vitro 31P NMR. The maximum specific growth rate and biomass yield obtained on glucose were twice as low as on sucrose. This was the result of low ATP levels being present in glucose-metabolizing cells, although the ATP production flux was as high as in sucrose-metabolizing cells due to a twofold increase of enzyme activities in both glycolytic pathways. Growth performance on glucose could be improved by adding fructose as an external electron acceptor, suggesting that the observed behavior is due to a redox imbalance causing energy starvation. PMID:17965151

  11. Genomic and genetic characterization of the bile stress response of probiotic Lactobacillus reuteri ATCC 55730.

    Science.gov (United States)

    Whitehead, Kristi; Versalovic, James; Roos, Stefan; Britton, Robert A

    2008-03-01

    Probiotic bacteria encounter various stresses after ingestion by the host, including exposure to the low pH in the stomach and bile in the small intestine. The probiotic microorganism Lactobacillus reuteri ATCC 55730 has previously been shown to survive in the human small intestine. To address how L. reuteri can resist bile stress, we performed microarray experiments to determine gene expression changes that occur when the organism is exposed to physiological concentrations of bile. A wide variety of genes that displayed differential expression in the presence of bile indicated that the cells were dealing with several types of stress, including cell envelope stress, protein denaturation, and DNA damage. Mutations in three genes were found to decrease the strain's ability to survive bile exposure: lr1864, a Clp chaperone; lr0085, a gene of unknown function; and lr1516, a putative esterase. Mutations in two genes that form an operon, lr1584 (a multidrug resistance transporter in the major facilitator superfamily) and lr1582 (unknown function), were found to impair the strain's ability to restart growth in the presence of bile. This study provides insight into the possible mechanisms that L. reuteri ATCC 55730 may use to survive and grow in the presence of bile in the small intestine. PMID:18245259

  12. Ferulic acid transformation into the main vanilla aroma compounds by Amycolatopsis sp. ATCC 39116.

    Science.gov (United States)

    Pérez-Rodríguez, Noelia; Oliveira, Ricardo Pinheiro de Souza; Agrasar, Ana María Torrado; Domínguez, José Manuel

    2016-02-01

    The wild strain Amycolatopsis sp. ATCC 39116 was explored in ferulic acid-based media to produce naturally the aroma components of the cured vanilla pod, namely vanillin,vanillic acid, and vanillyl alcohol. Other phenolic compounds(4-vinyl guaiacol, guaiacol, and protocatechuic acid) were also evaluated. The influence of medium composition,fermentation technology (batch or fed-batch), supplementation with vanillic acid, and inoculum concentration on ferulic acid biotransformation were evaluated. The results postulate the initial concentration of cell mass as the variable with the strongest impact on ferulic acid metabolization under the studied conditions. The highest amounts of vanillin and vanillic acid were achieved at intermediate values of cell mass.Vanillyl alcohol and protocatechuic acid were more closely linked to high cell mass concentrations. Conversely, 4-vinyl guaiacol reached its highest amount at the lowest amount of cell mass. Guaiacol was not detected in any case. Therefore,the initial cell concentration must be considered a critical parameter when using Amycolaptosis sp. ATCC 39116 for the production of vanillin and related compounds.

  13. Ferulic acid transformation into the main vanilla aroma compounds by Amycolatopsis sp. ATCC 39116.

    Science.gov (United States)

    Pérez-Rodríguez, Noelia; Oliveira, Ricardo Pinheiro de Souza; Agrasar, Ana María Torrado; Domínguez, José Manuel

    2016-02-01

    The wild strain Amycolatopsis sp. ATCC 39116 was explored in ferulic acid-based media to produce naturally the aroma components of the cured vanilla pod, namely vanillin,vanillic acid, and vanillyl alcohol. Other phenolic compounds(4-vinyl guaiacol, guaiacol, and protocatechuic acid) were also evaluated. The influence of medium composition,fermentation technology (batch or fed-batch), supplementation with vanillic acid, and inoculum concentration on ferulic acid biotransformation were evaluated. The results postulate the initial concentration of cell mass as the variable with the strongest impact on ferulic acid metabolization under the studied conditions. The highest amounts of vanillin and vanillic acid were achieved at intermediate values of cell mass.Vanillyl alcohol and protocatechuic acid were more closely linked to high cell mass concentrations. Conversely, 4-vinyl guaiacol reached its highest amount at the lowest amount of cell mass. Guaiacol was not detected in any case. Therefore,the initial cell concentration must be considered a critical parameter when using Amycolaptosis sp. ATCC 39116 for the production of vanillin and related compounds. PMID:26476645

  14. Cloning, expression and characterization of 3-hydroxyisobutyrate dehydrogenase from Pseudomonas denitrificans ATCC 13867.

    Directory of Open Access Journals (Sweden)

    Shengfang Zhou

    Full Text Available The gene encoding an NAD(+-dependent, 3-hydroxyisobutyrate dehydrogenase (3HIBDH-IV from Pseudomonas denitrificans ATCC 13867 was cloned and expressed in Escherichia coli BL 21 (DE3 and characterized to understand its physiological relevance in the degradation of 3-hydroxypropionic acid (3-HP. The deduced amino acid sequence showed high similarity to other 3-hydroxyisobutyrate dehydrogenase isozymes (3HIBDHs of P. denitrificans ATCC 13867. A comparison of 3HIBDH-IV with its relevant enzymes along with molecular docking studies suggested that Lys171, Asn175 and Gly123 are important for its catalytic function on 3-hydroxyacids. The recombinant 3HIBDH-IV was purified to homogeneity utilizing a Ni-NTA-HP resin column in high yield. 3HIBDH-IV was very specific to (S-3-hydroxyisobutyrate, but also catalyzed the oxidation of 3-HP to malonate semialdehyde. The specific activity and half-saturation constant (K m for 3-HP at 30°C and pH 9.0 were determined to be 17 U/mg protein and 1.0 mM, respectively. Heavy metals, such as Ag(+ and Hg(2+, completely inhibited the 3HIBDH-IV activity, whereas dithiothreitol, 2-mercaptoethanol and ethylenediaminetetraacetic acid increased its activity 1.5-1.8-fold. This paper reports the characteristics of 3HIBDH-IV as well as its probable role in 3-HP degradation.

  15. Transcriptomic analysis of (group I Clostridium botulinum ATCC 3502 cold shock response.

    Directory of Open Access Journals (Sweden)

    Elias Dahlsten

    Full Text Available Profound understanding of the mechanisms foodborne pathogenic bacteria utilize in adaptation to the environmental stress they encounter during food processing and storage is of paramount importance in design of control measures. Chill temperature is a central control measure applied in minimally processed foods; however, data on the mechanisms the foodborne pathogen Clostridium botulinum activates upon cold stress are scarce. Transcriptomic analysis on the C. botulinum ATCC 3502 strain upon temperature downshift from 37°C to 15°C was performed to identify the cold-responsive gene set of this organism. Significant up- or down-regulation of 16 and 11 genes, respectively, was observed 1 h after the cold shock. At 5 h after the temperature downshift, 199 and 210 genes were up- or down-regulated, respectively. Thus, the relatively small gene set affected initially indicated a targeted acute response to cold shock, whereas extensive metabolic remodeling appeared to take place after prolonged exposure to cold. Genes related to fatty acid biosynthesis, oxidative stress response, and iron uptake and storage were induced, in addition to mechanisms previously characterized as cold-tolerance related in bacteria. Furthermore, several uncharacterized DNA-binding transcriptional regulator-encoding genes were induced, suggesting involvement of novel regulatory mechanisms in the cold shock response of C. botulinum. The role of such regulators, CBO0477 and CBO0558A, in cold tolerance of C. botulinum ATCC 3502 was demonstrated by deteriorated growth of related mutants at 17°C.

  16. Levan fructotransferase from Arthrobacter oxydans J17-21 catalyzes the formation of the di-D-fructose dianhydride IV from levan.

    Science.gov (United States)

    Jang, Ki-Hyo; Ryu, Eun-Ja; Park, Buem-Seek; Song, Ki-Bang; Kang, Soon Ah; Kim, Chul Ho; Uhm, Tai-Boong; Park, Yong-Il; Rhee, Sang-Ki

    2003-04-23

    A new levan fructotransferase (LFTase) isolated from Arthrobacter oxydans J17-21 was characterized for the production of difructose dianhydride IV (DFA IV). LFTase was purified to apparent homogeneity by Q-Sepharose ion exchange chromatography, Mono-Q HR 5/5 column chromatography, and gel permeation chromatography. The enzyme had an apparent molecular mass of 54000 Da. The optimum pH for the enzyme-catalyzed reaction was pH 6.5, and the optimum temperature was observed at 45 degrees C. The LFTase was activated by the presence of CaCl(2) and EDTA-2Na but inhibited strongly by MnCl(2) and CuSO(4) at 1 mM and completely by FeSO(4) and Ag(2)SO(4) at 1 mM. A bacterial levan from Zymomonas mobilis was incubated with an LFTase; final conversion yield from the levan to DFA IV was 35%. Neither inulin, levanbiose, sucrose, dextran, nor starch was hydrolyzed by LFTase. DFA IV was very stable at acidic pH and high temperature, thus indicating that DFA IV may be suitable for the food industry and related areas. PMID:12696949

  17. Probing the Role of Two Critical Residues in Inulin Fructotransferase (DFA III-Producing) Thermostability from Arthrobacter sp. 161MFSha2.1.

    Science.gov (United States)

    Yu, Shuhuai; Wang, Xiao; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

    2016-08-10

    Inulin fructotransferase (IFTase) is an important enzyme that produces di-d-fructofuranose 1,2':2,3' dianhydride (DAF III), which is beneficial for human health. Present investigations mainly focus on screening and characterizing IFTase, including catalytic efficiency and thermostability, which are two important factors for enzymatic industrial applications. However, few reports aimed to improve these two characteristics based on the structure of IFTase. In this work, a structural model of IFTase (DFA III-producing) from Arthrobacter sp. 161MFSha2.1 was constructed through homology modeling. Analysis of this model reveals that two residues, Ser-309 and Ser-333, may play key roles in the structural stability. Therefore, the functions of the two residues were probed by site-directed mutagenesis combined with the Nano-DSC method and assays for residual activity. In contrast to other mutations, single mutation of serine 309 (or serine 333) to threonine did not decrease the enzymatic stability, whereas double mutation (serine 309 and serine 333 to threonine) can enhance thermostability (by approximately 5 °C). The probable mechanisms for this enhancement were investigated.

  18. Interaction mechanism between Arthrobacter oxydans and BaP-Cd%氧化节杆菌与苯并[a]芘-镉交互作用机理

    Institute of Scientific and Technical Information of China (English)

    邓军; 尹华; 叶锦韶; 彭辉; 秦华明; 龙焰; 何宝燕; 张娜

    2010-01-01

    利用HPLC、FTIR分析技术,研究了氧化节杆菌(Arthrobacter oxydans)对水体中苯并[a]芘(BaP)-镉(Cd)复合污染物交互作用机理.结果表明,BaP、Cd及BaP-Cd复合污染均对菌株的正常生长产生影响,其影响程度为:BaP-Cd>Cd>BaP;使用该菌投加到BaP、Cd浓度均为1 mg·L~(-1)的BaP-Cd复合污染无机盐溶液中,30 ℃、130 r·min~(-1),培养5 d后,体系BaP、Cd的残留量分别为0.39、0.65 mg·L~(-1);添加一定浓度的H_2O_2并保持体系呈偏碱性更适于该复合污染的处理.红外扫描分析显示,在微生物作用下,BaP的结构发生改变,菌株降解BaP和吸附Cd的过程主要与羟基、CC键、酰胺基团和C-H键有关.

  19. Characteristics of an organic solvent-tolerant β-fructofuranosidase from Arthrobacter arilaitensis NJEM01 and efficient synthesis of prebiotic kestose.

    Science.gov (United States)

    Chu, Jianlin; Wu, Xueming; Wu, Bin; Wang, Rui; He, Bingfang

    2014-06-18

    An organic solvent-tolerant β-fructofuranosidase (β-FFase) from Arthrobacter arilaitensis NJEM01 was purified, characterized, cloned, and overexpressed in Escherichia coli. The mature β-FFase contained 495 amino acid residues with an estimated molecular mass of 55 kDa. The purified β-FFase from strain NJEM01 was very stable in the buffer systems (pH 5.0-9.5) and showed high stability below 45 °C. Furthermore, the enzyme exhibited relatively high solvent stability in various aqueous organic mixtures and retained nearly 100% of its initial activity after incubation for 10 days in 20% (v/v) DMSO. In addition, the β-FFase exhibited high transfructosylation activity, synthesized prebiotic products of mainly 6-kestose (up to 476 g/L), and showed fructosyl receptor specificity to C-glucosyl flavone. A relatively high yield of FOS was achieved by the β-FFase from bacterium with a high concentration of sucrose. It made the β-FFase an exploitable biocatalyst for the production of glycosides of natural products and prebiotic kestose. PMID:24854707

  20. Cloning, nucleotide sequence and expression of a new L-N-carbamoylase gene from Arthrobacter aurescens DSM 3747 in E. coli.

    Science.gov (United States)

    Wilms, B; Wiese, A; Syldatk, C; Mattes, R; Altenbuchner, J; Pietzsch, M

    1999-02-19

    An L-N-carbamoyl amino acid amidohydrolase (L-N-carbamoylase) from Arthrobacter aurescens DSM 3747 was cloned in E. coli and the nucleotide sequence was determined. After expression of the gene in E. coli the enzyme was purified to homogeneity and characterized. The enzyme was shown to be strictly L-specific and exhibited the highest activity in the hydrolysis of beta-aryl substituted N alpha-carbamoyl-alanines as e.g. N-carbamoyl-tryptophan. Carbamoyl derivatives of beta-alanine and charged aliphatic amino acids were not accepted as substrates. The N-carbamoylase of A. aurescens DSM 3747 differs from all known enzymes with respect to its substrate specificity although amino acid sequence identity scores of 35-38% to other N-carbamoylases have been detected. The enzyme consists of two subunits of 44,000 Da, and has an isoelectric point of 4.3. The optima of temperature and pH were determined to be 50 degrees C and pH 8.5 respectively. At 37 degrees C the enzyme was completely stable for several days. PMID:10194852

  1. Inactivation of Listeria monocytogenes ATCC 7644 on fresh-cut tomato using nisin in combinations with organic salts

    OpenAIRE

    Oladunjoye, Adebola O.; Singh, Suren; Ijabadeniyi, Oluwatosin A.

    2016-01-01

    The inhibition of Listeria monocytogenes ATCC 7644 on fresh-cut tomato was investigated using nisin alone, and in combinations with organic salts. Nisin at a concentration of 5000 UI/mL was introduced alone or in combination with an organic salt (sodium citrate or sodium acetate each at 3 and 5 g/100 mL each) on fresh-cut tomato previously inoculated with 108 CFU/mL of L. monocytogenes ATCC 7644. Chlorine at 200 ppm was used as a control. The inoculated samples were incubated at different tem...

  2. Suitability of Lactococcus lactis subsp lactis ATCC 11454 as a protective culture for lightly preserved fish products

    DEFF Research Database (Denmark)

    Wessels, Stephen Wallace; Huss, Hans Henrik

    1996-01-01

    This study is part of strategy to control the human pathogen Listeria monocytogenes in lightly preserved fish products by using food-grade lactic acid bacteria. When the nisin-producing Lactococcus lactis subsp lactis ATCC 11454 was cultured in the same vessel as L-monocytogenes Scott A in brain......-heart infusion broth (BHI) at 30-degrees C, the pathogen declined from 5x10(5) to fewer than 5 cfu ml(-1) within 31 h. The effect was not due to lactic acid inhibition. Growth and nisin production by L- lactis ATCC 11454 were investigated under the conditions of temperature and salt used for light preservation...

  3. High-Quality Draft Genome Sequence of Aneurinibacillus migulanus ATCC 9999T (DSM 2895), a Gramicidin S-Producing Bacterium Isolated from Garden Soil.

    Science.gov (United States)

    Wang, Jie-Ping; Liu, Bo; Liu, Guo-Hong; Ge, Ci-Bin; Xiao, Rong-Feng; Zheng, Xue-Fang; Shi, Huai

    2015-01-01

    Aneurinibacillus migulanus ATCC 9999(T) (DSM 2895) is a Gram-positive, round-spore-forming, and gramicidin S-producing bacterium. Here, we report the 6.35-Mb high-quality draft genome sequence of A. migulanus ATCC 9999(T), which will provide useful information for the genomic taxonomy and phylogenomics of Bacillus-like bacteria. PMID:26494674

  4. Isolation and partial characterization of protease from Pseudomonas aeruginosa ATCC 27853

    Directory of Open Access Journals (Sweden)

    LIDIJA IZRAEL-ŽIVKOVIĆ

    2010-08-01

    Full Text Available Enzymatic characteristics of a protease from a medically important, referent strain of Pseudomonas aeruginosa ATCC 27853 were determined. According to sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE, and gel filtration, it was estimated that the molecular mass of the purified enzyme was about 15 kDa. Other enzymatic properties were found to be: pH optimum 7.1, pH stability between 6.5 and 10; temperature optimum around 60 °C while the enzyme was stable at 60 °C for 30 min. Inhibition of the enzyme was observed with metal chelators, such as EDTA and 1,10-phenanthroline, suggesting that the protease is a metalloenzyme. Furthermore, the enzyme contains one mole of zinc ion per mole of enzyme. The protease was stable in the presence of different organic solvents, which enables its potential use for the synthesis of peptides.

  5. Sensibility of salmonella typhimurium (atcc 0626) to gamma radiation in minced breast chicken

    International Nuclear Information System (INIS)

    Samples of raw minced breast chicken were inoculated with 10 7 cfu/ml of salmonella typhimurium (atcc 0626) and submitted to gamma radiation.The cobalt-60 source was a gamma beam 650, with a dose rate of 929 Gy/h.Samples were irradiated at room temperature (25-27 0c) with doses of 2, 4, 6 and 8 KGy. After irradiation the samples, including the control, were kept under refrigeration condition (5 0c) and at day 1, 7, 14, 21 and 28 of storage, were analyzed for s. Typhimurium presence. The 2 KGy dose inhibited the bacteria growth at the 21 st day; the dose of 4 KGy was effective at the 7 t h day. The doses of 6 KGy and 8 KGy destroyed the bacteria just after the irradiation process

  6. Sensibility Of Salmonella typhimurium (ATCC 0626) to gamma radiation in minced breast Chicken

    International Nuclear Information System (INIS)

    Samples of raw minced breast chicken were inoculated with 10 7 CFU/mL of Salmonella typhimurium (ATCC 0626) and submitted to gamma radiation. The Cobalt-60 source was a Gamma beam 650, with a dose rate of 929 Gy/h. Samples were irradiated at room temperature (25-270C) with doses of 2, 4, 6 and 8 kGy. After irradiation the samples, including the control, were kept under refrigeration condition (5 0C) and at day 1, 7, 14, 21 and 28 of storage, were analyzed for S. typhimurium presence. The 2 kGy dose inhibited the bacteria growth at the 21 st day; the dose of 4 kGy was effective at the 7 t h day. The doses of 6 kGy and 8 kGy destroyed the bacteria just after the irradiation process

  7. Exploration of geosmin synthase from Streptomyces peucetius ATCC 27952 by deletion of doxorubicin biosynthetic gene cluster.

    Science.gov (United States)

    Singh, Bijay; Oh, Tae-Jin; Sohng, Jae Kyung

    2009-10-01

    Thorough investigation of Streptomyces peucetius ATCC 27952 genome revealed a sesquiterpene synthase, named spterp13, which encodes a putative protein of 732 amino acids with significant similarity to S. avermitilis MA-4680 (SAV2163, GeoA) and S. coelicolor A3(2) (SCO6073). The proteins encoded by SAV2163 and SCO6073 produce geosmin in the respective strains. However, the spterp13 gene seemed to be silent in S. peucetius. Deletion of the doxorubicin gene cluster from S. peucetius resulted in increased cell growth rate along with detectable production of geosmin. When we over expressed the spterp13 gene in S. peucetius DM07 under the control of an ermE* promoter, 2.4 +/- 0.4-fold enhanced production of geosmin was observed.

  8. Effects of Salt Stress on Carbohydrate Metabolism of Lactobacillus plantarum ATCC 14917.

    Science.gov (United States)

    Wang, Pingping; Wu, Zhen; Wu, Jing; Pan, Daodong; Zeng, Xiaoqun; Cheng, Kemeng

    2016-10-01

    Lactic acid bacteria are widely used in fermented foods, especially cheese products. In this study, we observed the salt tolerance of Lactobacillus plantarum ATCC 14917 after exposure to different concentrations of NaCl in MRS medium. Quantitative proteomic profiles using two-dimensional electrophoresis identified 384 proteins, of which 26 were upregulated and 31 downregulated. Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry was then used to identify 11 proteins, of which three were linked to carbohydrate metabolism. The downregulation of carbamoyl phosphate synthase in carbohydrate metabolism revealed a bacterial regulation mechanism to save energy in order to survive during the salt tolerance. Other proteins were found involved in transcription-translation processes, fatty acid biosynthesis, and the primary metabolic process. PMID:27342422

  9. Lactobacillus reuteri ATCC 55730 and L22 display probiotic potential in vitro and protect against Salmonella-induced pullorum disease in a chick model of infection.

    Science.gov (United States)

    Zhang, Dexian; Li, Rui; Li, Jichang

    2012-08-01

    Lactobacillus reuteri ATCC 55730 (L. reuteri ATCC 55730) and L. reuteri L22 were studied for their probiotic potential. These two strains were able to produce an antimicrobial substance, termed reuterin, the maximum production of reuterin by these two strains was detected in the late logarithmic growth phase (16 h in MRS and 20 h in LB broths). These two strains could significantly reduce the growth of Salmonella pullorum ATCC 9120 in MRS broth, L. reuteri ATCC 55730 with a reduction of 48.2±4.15% (in 5 log) and 89.7±2.59% (in 4 log) respectively, at the same time, L. reuteri L22 was 69.4±3.48% (in 5 log) and 80.4±3.22% respectively. L. reuteri ATCC 55730 was active against the majority of the pathogenic species, including S. pullorum ATCC 9120 and Escherichia coli O(78), while L. reuteri L22 was not as effective as L. reuteri ATCC 55730. The two potential strains were found to survive variably at pH 2.5 and were unaffected by bile salts, while neither of the strains was haemolytic. Moreover, L. reuteri ATCC 55730 exhibited variable susceptibility towards commonly used antibiotics; but L. reuteri L22 showed resistant to most antibiotics in this study. L. reuteri ATCC 55730 consequently was found to significantly increase survival rate in a Salmonella-induced pullorum disease model in chick. To conclude, strain L. reuteri ATCC 55730 possesses desirable probiotic properties, such as antimicrobial activity and immunomodulation in vitro, which were confirmed in vivo by the use of animal models. PMID:21764090

  10. Quantitative analysis of population heterogeneity of the adaptive salt stress response and growth capacity of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Besten, den H.M.W.; Ingham, C.J.; Hylckama Vlieg, van J.E.T.; Beerthuyzen, M.M.; Zwietering, M.H.; Abee, T.

    2007-01-01

    Bacterial populations can display heterogeneity with respect to both the adaptive stress response and growth capacity of individual cells. The growth dynamics of Bacillus cereus ATCC 14579 during mild and severe salt stress exposure were investigated for the population as a whole in liquid culture.

  11. Construction of expression vectors for metabolic engineering of the vanillin-producing actinomycete Amycolatopsis sp. ATCC 39116.

    Science.gov (United States)

    Fleige, Christian; Steinbüchel, Alexander

    2014-01-01

    Amycolatopsis sp. ATCC 39116 is able to synthesize the important flavoring agent vanillin from cheap natural substrates. The bacterium is therefore of great interest for the industry and used for the fermentative production of vanillin. In order to improve the production of natural vanillin with Amycolatopsis sp. ATCC 39116, the strain has been genetically engineered to optimize the metabolic flux towards the desired product. Extensive metabolic engineering was hitherto hampered, due to the lack of genetic tools like functional promoters and expression vectors. In this study, we report the establishment of a plasmid-based gene expression system for Amycolatopsis sp. ATCC 39116 that allows a further manipulation of the genotype. Four new Escherichia coli-Amycolatopsis shuttle vectors harboring different promoter elements were constructed, and the functionality of these regulatory elements was proven by the expression of the reporter gene gusA, encoding a β-glucuronidase. Glucuronidase activity was detected in all plasmid-harboring strains, and remarkable differences in the expression strength of the reporter gene depending on the used promoter were observed. The new expression vectors will promote the further genetic engineering of Amycolatopsis sp. ATCC 39116 to get insight into the metabolic network and to improve the strain for a more efficient industrial use. PMID:24743982

  12. Genome Sequence of Corynebacterium glutamicum ATCC 14067, Which Provides Insight into Amino Acid Biosynthesis in Coryneform Bacteria

    OpenAIRE

    Lv, Yangyong; Liao, Juanjun; Wu, Zhanhong; Han, Shuangyan; Lin, Ying; Zheng, Suiping

    2012-01-01

    We report the genome sequence of Corynebacterium glutamicum ATCC 14067 (once named Brevibacterium flavum), which is useful for taxonomy research and further molecular breeding in amino acid production. Preliminary comparison with those of the reported coryneform strains revealed some notable differences that might be related to the difficulties in molecular manipulation.

  13. Complete Genome Sequence of Gluconacetobacter hansenii Strain NQ5 (ATCC 53582), an Efficient Producer of Bacterial Cellulose.

    Science.gov (United States)

    Pfeffer, Sarah; Mehta, Kalpa; Brown, R Malcolm

    2016-08-11

    This study reports the release of the complete nucleotide sequence of Gluconacetobacter hansenii strain NQ5 (ATCC 53582). This strain was isolated by R. Malcolm Brown, Jr. in a sugar mill in North Queensland, Australia, and is an efficient producer of bacterial cellulose. The elucidation of the genome will contribute to the study of the molecular mechanisms necessary for cellulose biosynthesis.

  14. Direct-Imaging-Based Quantification of Bacillus cereus ATCC 14579 Population Heterogeneity at a Low Incubation Temperature

    NARCIS (Netherlands)

    Besten, den H.M.W.; Garcia, D.; Moezelaar, R.; Zwietering, M.H.; Abee, T.

    2010-01-01

    Bacillus cereus ATCC 14579 was cultured in microcolonies on Anopore strips near its minimum growth temperature to directly image and quantify its population heterogeneity at an abusive refrigeration temperature. Eleven percent of the microcolonies failed to grow during low-temperature incubation, an

  15. Fe(III) stimulates 3-methylindole and 4-methylphenol production in swine lagoon enrichments and Clostridium scatologenes ATCC 25775

    Science.gov (United States)

    Aims: To determine the effects of anaerobic electron acceptors on 3-methylindole (3-MI) and 4-methylphenol (4-MP) production in swine waste lagoon enrichments and Clostridium scatologenes ATCC 25775. Methods and Results: Swine waste lagoon sediment was incubated anaerobically in tryptone-yeast ext...

  16. Effect of Lactobacillus brevis ATCC 8287 as a feeding supplement on the performance and immune function of piglets

    Science.gov (United States)

    Lactobacillus brevis ATCC 8287, a surface (S-layer) strain, possesses a variety of functional properties that make it both a potential probiotic and a good vaccine vector candidate. With this in mind, our aim was to study the survival of L. brevis in the porcine gut and investigate the effect of th...

  17. Functional analysis of the gene cluster involved in production of the bacteriocin circularin A by Clostridium beijerinckii ATCC 25752

    NARCIS (Netherlands)

    Kemperman, R; Jonker, M; Nauta, A; Kuipers, OP; Kok, J

    2003-01-01

    A region of 12 kb flanking the structural gene of the cyclic antibacterial peptide circularin A of Clostridium beijerinckii ATCC 25752 was sequenced, and the putative proteins involved in the production and secretion of circularin A were identified. The genes are tightly organized in overlapping ope

  18. Complete Genome Sequence of Gluconacetobacter hansenii Strain NQ5 (ATCC 53582), an Efficient Producer of Bacterial Cellulose.

    Science.gov (United States)

    Pfeffer, Sarah; Mehta, Kalpa; Brown, R Malcolm

    2016-01-01

    This study reports the release of the complete nucleotide sequence of Gluconacetobacter hansenii strain NQ5 (ATCC 53582). This strain was isolated by R. Malcolm Brown, Jr. in a sugar mill in North Queensland, Australia, and is an efficient producer of bacterial cellulose. The elucidation of the genome will contribute to the study of the molecular mechanisms necessary for cellulose biosynthesis. PMID:27516505

  19. Growth and sporulation of Bacillus cereus ATCC 14579 under defined conditions: temporal expression of genes for key sigma factors

    NARCIS (Netherlands)

    Vries, de Y.P.; Hornstra, L.M.; Vos, de W.M.; Abee, T.

    2004-01-01

    An airlift fermentor system allowing precise regulation of pH and aeration combined with a chemically defined medium was used to study growth and sporulation of Bacillus cereus ATCC 14579. Sporulation was complete and synchronous. Expression of sigA, sigB, sigF, and sigG was monitored with real-time

  20. Production of the glycopeptide antibiotic A40926 by Nonomuraea sp ATCC 39727: influence of medium composition in batch fermentation

    DEFF Research Database (Denmark)

    Gunnarsson, Nina; Bruheim, Per; Nielsen, Jens

    2003-01-01

    Nonomuraea sp. ATCC 39727 is a novel actinomycete species and the producer of A40926, a glycopeptide antibiotic structurally similar to teichoplanin. In the present study, a defined minimal medium was designed for Nonomuraea fermentation. The influence of initial phosphate, glucose and ammonium...

  1. Genome sequence alterations detected upon passage of Burkholderia mallei ATCC 23344 in culture and in mammalian hosts

    Directory of Open Access Journals (Sweden)

    Yu Yan

    2006-09-01

    Full Text Available Abstract Background More than 12,000 simple sequence repeats (SSRs have been identified in the genome of Burkholderia mallei ATCC 23344. As a demonstrated mechanism of phase variation in other pathogenic bacteria, these may function as mutable loci leading to altered protein expression or structure variation. To determine if such alterations are occurring in vivo, the genomes of various single-colony passaged B. mallei ATCC 23344 isolates, one from each source, were sequenced from culture, a mouse, a horse, and two isolates from a single human patient, and the sequence compared to the published B. mallei ATCC 23344 genome sequence. Results Forty-nine insertions and deletions (indels were detected at SSRs in the five passaged strains, a majority of which (67.3% were located within noncoding areas, suggesting that such regions are more tolerant of sequence alterations. Expression profiling of the two human passaged isolates compared to the strain before passage revealed alterations in the mRNA levels of multiple genes when grown in culture. Conclusion These data support the notion that genome variability upon passage is a feature of B. mallei ATCC23344, and that within a host B. mallei generates a diverse population of clones that accumulate genome sequence variation at SSR and other loci.

  2. Characterization of inosine monophosphate dehydrogenase from Staphylococcus aureus ATCC12600 and its involvement in biofilm formation

    Directory of Open Access Journals (Sweden)

    S. Yeswanth

    2013-10-01

    Full Text Available Background: In Staphylococcus aureus purine metabolism plays a crucial role in the formation of biofilm which is a key pathogenic factor. The present study is aimed in the characterization of inosine monophosphate dehydrogenase (IMPDH from Staphylococcus aureus ATCC 12600. Methods: IMPDH gene was amplified using primers designed from IMPDH gene sequence of S. aureus reported in the database. Then polymerase chain reaction (PCR product was cloned in the Sma I site of M13mp18 and expressed in Escherichia coli JM109. The recombinant IMPDH (rIMPDH was overexpressed with 1 mM isopropyl beta-D-1- thiogalactopyranoside (IPTG; Michaelis constant (Km, maximum enzyme velocity (Vmax and catalytic constant (Kcat of expressed IMPDH were determined. Results: The enzyme kinetics of IMPDH grown under aerobic conditions showed a Km of 43.71±1.56 µM, Vmax of 0.247±0.84/µM/mg/min and Kcat of 2.74±0.015/min while in anaerobic conditions the kinetics showed Km of 42.81±3.154/ µM, Vmax of 0.378±0.036 µM/mg/min and Kcat of 4.78±0.021 /min, indicating elevated levels of IMPDH activity under anaerobic conditions. Three-folds increased activity in the presence of 1 mM adenosine triphosphate (ATP correlated with biofilm formation. The kinetics of pure rIMPDH were close to the native IMPDH of S. aureus ATCC12600 and the enzyme showed single band in sodium dodecyl sulphate polyacrylamide gel electrophoresis with a molecular weight of 53 KDa. Conclusions: Elevated activity of IMPDH was observed in S. aureus grown under anaerobic conditions and this was correlated with the biofilm formation indicating the linkage between purine metabolism and pathogenesis.

  3. Meta-analysis: Lactobacillus reuteri strain DSM 17938 (and the original strain ATCC 55730) for treating acute gastroenteritis in children.

    Science.gov (United States)

    Szajewska, H; Urbańska, M; Chmielewska, A; Weizman, Z; Shamir, R

    2014-09-01

    Lactobacillus reuteri ATCC 55730 has been shown to provide a moderate clinical effect in the treatment of acute gastroenteritis (AGE) in children. However, as the L. reuteri ATCC 55730 strain was found to carry potentially transferable resistance traits for tetracycline and lincomycin, it was replaced by a new strain, L. reuteri DSM 17938, without unwanted plasmid-borne antibiotic resistance. Bioequivalence of the two strains has been suggested. We aimed to systematically evaluate data on the effectiveness of L. reuteri DSM 17938 and the original strain, L. reuteri ATCC 55730, in the treatment of AGE in children. The Cochrane Library, MEDLINE, and EMBASE databases, reference lists, and abstract books of major scientific meetings were searched in August 2013, with no language restrictions, for relevant randomised controlled trials (RCTs). Two RCTs (n=196) that evaluated L. reuteri DSM 17938 and three RCTs (n=156) that evaluated L. reuteri ATCC 55730, which involved hospitalised children aged 3 to 60 months, met the inclusion criteria. Compared with placebo or no treatment, DSM 17938 significantly reduced the duration of diarrhoea (mean difference -32 h, 95% confidence interval (CI): -41 to -24) and increased the chance of cure on day 3 (relative risk: 3.5, 95% CI: 1.2 to 10.8, random effects model). Similar results were obtained with the original strain, L. reuteri ATCC 55730. In conclusion, in hospitalised children, use of both strains of L. reuteri reduced the duration of diarrhoea, and more children were cured within 3 days. Data from outpatients and countryspecific cost-effectiveness analyses are needed. Given the limited data and the methodological limitations of the included trials, the evidence should be viewed with caution. PMID:24463209

  4. In situ hydrogen, acetone, butanol, ethanol and microdiesel production by Clostridium acetobutylicum ATCC 824 from oleaginous fungal biomass.

    Science.gov (United States)

    Hassan, Elhagag Ahmed; Abd-Alla, Mohamed Hemida; Bagy, Magdy Mohamed Khalil; Morsy, Fatthy Mohamed

    2015-08-01

    An in situ batch fermentation technique was employed for biohydrogen, acetone, butanol, ethanol and microdiesel production from oleaginous fungal biomass using the anaerobic fermentative bacterium Clostridium acetobutylicum ATCC 824. Oleaginous fungal Cunninghamella echinulata biomass which has ability to accumulate up to 71% cellular lipid was used as the substrate carbon source. The maximum cumulative hydrogen by C. acetobutylicum ATCC 824 from crude C. echinulata biomass was 260 ml H2 l(-1), hydrogen production efficiency was 0.32 mol H2 mole(-1) glucose and the hydrogen production rate was 5.2 ml H2 h(-1). Subsequently, the produced acids (acetic and butyric acids) during acidogenesis phase are re-utilized by ABE-producing clostridia and converted into acetone, butanol, and ethanol. The total ABE produced by C. acetobutylicum ATCC 824 during batch fermentation was 3.6 g l(-1) from crude fungal biomass including acetone (1.05 g l(-1)), butanol (2.19 g l(-1)) and ethanol (0.36 g l(-1)). C. acetobutylicum ATCC 824 has ability to produce lipolytic enzymes with a specific activity 5.59 U/mg protein to hydrolyze ester containing substrates. The lipolytic potential of C. acetobutylicum ATCC 824 was used as a biocatalyst for a lipase transesterification process using the produced ethanol from ABE fermentation for microdiesel production. The fatty acid ethyl esters (microdiesel) generated from the lipase transesterification of crude C. echinulata dry mass was analyzed by GC/MS as 15.4% of total FAEEs. The gross energy content of biohydrogen, acetone, butanol, ethanol and biodiesel generated through C. acetobutylicum fermentation from crude C. echinulata dry mass was 3113.14 kJ mol(-1). These results suggest a possibility of integrating biohydrogen, acetone, butanol and ethanol production technology by C. acetobutylicum with microdiesel production from crude C. echinulata dry mass and therefore improve the feasibility and commercialization of bioenergy production.

  5. In Silico Approach to Support that p-Nitrophenol Monooxygenase from Arthrobacter sp. Strain JS443 Catalyzes the Initial Two Sequential Monooxygenations.

    Science.gov (United States)

    Kallubai, Monika; Amineni, Umamaheswari; Mallavarapu, Megharaj; Kadiyala, Venkateswarlu

    2015-06-01

    p-Nitrophenol (PNP), used primarily for manufacturing pesticides and dyes, has been recognized as a priority environmental pollutant. It is therefore important to reduce the input of this toxicant into the environment and to establish approaches for its removal from the contaminated sites. PNP monooxygenase, a novel enzyme from Gram-positive bacteria like Arthrobacter sp. and Bacillus sp., that comprises two components, a flavoprotein reductase and an oxygenase, catalyzes the initial two sequential monooxygenations to convert PNP to trihydroxybenzene. Accurate and reliable prediction of this enzyme-substrate interactions and binding affinity are of vital importance in understanding these catalytic mechanisms of the two sequential reactions. As crystal structure of the enzyme has not yet been published, we built a homology model for PNP monooxygenase using crystallized chlorophenol 4-monooxygenase from Burkholderia cepacia AC1100 (3HWC) as the template. The model was assessed for its reliability using PROCHECK, ERRAT and ProSA. Molecular docking of the physiological substrates, PNP and 4-nitrocatechol (4-NC), was carried out using Glide v5.7 implemented in Maestro v9.2, and the binding energies were calculated to substantiate the prediction. Docking complexes formed by molecular level interactions of PNP monooxygenase-PNP/4-NC without or with the cofactors, FAD and NADH, showed good correlation with the established experimental evidence that the two-component PNP monooxygenase catalyzes both the hydroxylation of PNP and the oxidative release of nitrite from 4-NC in B. sphaericus JS905. Furthermore, molecular dynamics simulations performed for docking complexes using Desmond v3.0 showed stable nature of the interactions as well.

  6. Identification of a Novel Di-D-Fructofuranose 1,2’:2,3’ Dianhydride (DFA III) Hydrolysis Enzyme from Arthrobacter aurescens SK8.001

    Science.gov (United States)

    Yu, Shuhuai; Wang, Xiao; Zhang, Tao; Stressler, Timo; Fischer, Lutz; Jiang, Bo; Mu, Wanmeng

    2015-01-01

    Previously, a di-D-fructofuranose 1,2’:2,3’ dianhydride (DFA III)-producing strain, Arthrobacter aurescens SK8.001, was isolated from soil, and the gene cloning and characterization of the DFA III-forming enzyme was studied. In this study, a DFA III hydrolysis enzyme (DFA IIIase)-encoding gene was obtained from the same strain, and the DFA IIIase gene was cloned and expressed in Escherichia coli. The SDS-PAGE and gel filtration results indicated that the purified enzyme was a homotrimer holoenzyme of 145 kDa composed of subunits of 49 kDa. The enzyme displayed the highest catalytic activity for DFA III at pH 5.5 and 55°C, with specific activity of 232 U mg-1. Km and Vmax for DFA III were 30.7 ± 4.3 mM and 1.2 ± 0.1 mM min-1, respectively. Interestingly, DFA III-forming enzymes and DFA IIIases are highly homologous in amino acid sequence. The molecular modeling and docking of DFA IIIase were first studied, using DFA III-forming enzyme from Bacillus sp. snu-7 as a template. It was suggested that A. aurescens DFA IIIase shared a similar three-dimensional structure with the reported DFA III-forming enzyme from Bacillus sp. snu-7. Furthermore, their catalytic sites may occupy the same position on the proteins. Based on molecular docking analysis and site-directed mutagenesis, it was shown that D207 and E218 were two potential critical residues for the catalysis of A. aurescens DFA IIIase. PMID:26555784

  7. Arthrobacter aurescens TC1 Atrazine Catabolism Genes trzN, atzB, and atzC Are Linked on a 160-Kilobase Region and Are Functional in Escherichia coli

    OpenAIRE

    Sajjaphan, Kannika; Shapir, Nir; Wackett, Lawrence P; Palmer, Michael; Blackmon, Barbara; Tomkins, Jeff; Sadowsky, Michael J.

    2004-01-01

    Arthrobacter aurescens strain TC1 metabolizes atrazine to cyanuric acid via TrzN, AtzB, and AtzC. The complete sequence of a 160-kb bacterial artificial chromosome clone indicated that trzN, atzB, and atzC are linked on the A. aurescens genome. TrzN, AtzB, and AtzC were shown to be functional in Escherichia coli. Hybridization studies localized trzN, atzB, and atzC to a 380-kb plasmid in A. aurescens strain TC1.

  8. Structural analysis of Clostridium acetobutylicum ATCC 824 glycoside hydrolase from CAZy family GH105

    Energy Technology Data Exchange (ETDEWEB)

    Germane, Katherine L., E-mail: katherine.germane.civ@mail.mil [Oak Ridge Associated Universities, 4692 Millennium Drive, Suite 101, Belcamp, MD 21017 (United States); Servinsky, Matthew D. [US Army Research Laboratory, 2800 Powder Mill Road, Adelphi, MD 20783 (United States); Gerlach, Elliot S. [Federal Staffing Resources, 2200 Somerville Road, Annapolis, MD 21401 (United States); Sund, Christian J. [US Army Research Laboratory, 2800 Powder Mill Road, Adelphi, MD 20783 (United States); Hurley, Margaret M., E-mail: katherine.germane.civ@mail.mil [US Army Research Laboratory, 4600 Deer Creek Loop, Aberdeen Proving Ground, MD 21005 (United States); Oak Ridge Associated Universities, 4692 Millennium Drive, Suite 101, Belcamp, MD 21017 (United States)

    2015-07-29

    The crystal structure of the protein product of the C. acetobutylicum ATCC 824 gene CA-C0359 is structurally similar to YteR, an unsaturated rhamnogalacturonyl hydrolase from B. subtilis strain 168. Substrate modeling and electrostatic studies of the active site of the structure of CA-C0359 suggests that the protein can now be considered to be part of CAZy glycoside hydrolase family 105. Clostridium acetobutylicum ATCC 824 gene CA-C0359 encodes a putative unsaturated rhamnogalacturonyl hydrolase (URH) with distant amino-acid sequence homology to YteR of Bacillus subtilis strain 168. YteR, like other URHs, has core structural homology to unsaturated glucuronyl hydrolases, but hydrolyzes the unsaturated disaccharide derivative of rhamnogalacturonan I. The crystal structure of the recombinant CA-C0359 protein was solved to 1.6 Å resolution by molecular replacement using the phase information of the previously reported structure of YteR (PDB entry (http://scripts.iucr.org/cgi-bin/cr.cgi?rm)) from Bacillus subtilis strain 168. The YteR-like protein is a six-α-hairpin barrel with two β-sheet strands and a small helix overlaying the end of the hairpins next to the active site. The protein has low primary protein sequence identity to YteR but is structurally similar. The two tertiary structures align with a root-mean-square deviation of 1.4 Å and contain a highly conserved active pocket. There is a conserved aspartic acid residue in both structures, which has been shown to be important for hydration of the C=C bond during the release of unsaturated galacturonic acid by YteR. A surface electrostatic potential comparison of CA-C0359 and proteins from CAZy families GH88 and GH105 reveals the make-up of the active site to be a combination of the unsaturated rhamnogalacturonyl hydrolase and the unsaturated glucuronyl hydrolase from Bacillus subtilis strain 168. Structural and electrostatic comparisons suggests that the protein may have a slightly different substrate

  9. Actinoplanes teichomyceticus ATCC 31121 as a cell factory for producing teicoplanin

    Directory of Open Access Journals (Sweden)

    Taurino Carlo

    2011-10-01

    Full Text Available Abstract Background Teicoplanin is a glycopeptide antibiotic used clinically in Europe and in Japan for the treatment of multi-resistant Gram-positive infections. It is produced by fermenting Actinoplanes teichomyceticus. The pharmaceutically active principle is teicoplanin A2, a complex of compounds designated T-A2-1-A2-5 differing in the length and branching of the fatty acid moiety linked to the glucosamine residue on the heptapeptide scaffold. According to European and Japanese Pharmacopoeia, components of the drug must be reproduced in fixed amounts to be authorized for clinical use. Results We report our studies on optimizing the fermentation process to produce teicoplanin A2 in A. teichomyceticus ATCC 31121. Robustness of the process was assessed on scales from a miniaturized deep-well microtiter system to flasks and 3-L bioreactor fermenters. The production of individual factors T-A2-1-A2-5 was modulated by adding suitable precursors to the cultivation medium. Specific production of T-A2-1, characterized by a linear C10:1 acyl moiety, is enhanced by adding methyl linoleate, trilinoleate, and crude oils such as corn and cottonseed oils. Accumulation of T-A2-3, characterized by a linear C10:0 acyl chain, is stimulated by adding methyl oleate, trioleate, and oils such as olive and lard oils. Percentages of T-A2-2, T-A2-4, and, T-A2-5 bearing the iso-C10:0, anteiso-C11:0, and iso-C11:0 acyl moieties, respectively, are significantly increased by adding precursor amino acids L-valine, L-isoleucine, and L-leucine. Along with the stimulatory effect on specific complex components, fatty acid esters, oils, and amino acids (with the exception of L-valine inhibit total antibiotic productivity overall. By adding industrial oils to medium containing L-valine the total production is comparable, giving unusual complex compositions. Conclusions Since the cost and the quality of teicoplanin production depend mainly on the fermentation process, we

  10. Development of a potential functional food prepared with pigeon pea (Cajanus cajan), oats and Lactobacillus reuteri ATCC 55730.

    Science.gov (United States)

    Barboza, Yasmina; Márquez, Enrique; Parra, Katynna; Piñero, M Patricia; Medina, Luis M

    2012-11-01

    The purpose of this study was to investigate the survival of Lactobacillus reuteri ATCC 55730 in creams, prepared with pigeon peas and oat. Products were analysed to determine their content of protein, fibre, fat, carbohydrates and degree of likeness. Viable numbers of L. reuteri and pH were determined after 1, 7, 14, 21 and 28 days of storage at 4°C. Results showed significant differences (P 0.05) were found on sensory quality between control and creams with L. reuteri. After 28 days, the cell viability was above 7 log cfu/g in all creams. L. reuteri ATCC 55730 had the highest viability in cream with 40% pigeon pea and 20% oat (8.16 log cfu/g). In conclusion, due to its acceptability and highly nutritious value, the product could be used so as to support the growth of L. reuteri. PMID:22533458

  11. Listeria ivanovii ATCC 19119 strain behaviour is modulated by iron and acid stress.

    Science.gov (United States)

    Longhi, Catia; Ammendolia, Maria Grazia; Conte, Maria Pia; Seganti, Lucilla; Iosi, Francesca; Superti, Fabiana

    2014-09-01

    It has been suggested that the rarity of human listeriosis due to Listeria ivanovii reflects not only host tropism factors but also the rare occurrence of this species in the environment, compared with Listeria monocytogenes. In the present study we evaluate the effects on the reference strain L. ivanovii ATCC 19119 behaviour of two combined stresses, low iron availability and acid environment, that bacteria can encounter in the passage from saprophytic life to the host. In these conditions, L. ivanovii evidenced a different behaviour compared to L. monocytogenes exposed to similar conditions. L. ivanovii was not able to mount an acid tolerance response (ATR) even if, upon entry into the stationary phase in iron-loaded medium, growth phase-dependent acid resistance (AR) was evidenced. Moreover, bacteria grown in iron excess and acidic pH showed the higher invasion value in Caco-2 cells, even though it was not able to efficiently multiply. On the contrary, low iron and acidic conditions improved invasion ability in amniotic WISH cells.

  12. Incorporation of cholesterol into the cellular membrane of Lactobacillus acidophilus ATCC 43121

    International Nuclear Information System (INIS)

    Cholesterol that was assimilated by Lactobacillus acidophilus ATCC 43121 was not metabolically degraded; most of it was recovered with the cells. Cells that were grown in the presence of cholesterol micelles and bile salts were more resistant to lysis by sonication than were those grown in their absence, suggesting a possible alteration of the cell wall or membrane. Cholesterol assimilation occurred during growth at pH 6.0 as well as during growth without pH control. Part of the cholesterol that was assimilated by cells was recovered in the membrane fractions of cells grown under both conditions. There was no difference in the amount taken up from cholesterol micelles that were prepared using dioleoyl L-alpha-phosphatidylcholine or distearoyl L-alpha-phosphatidylcholine. Thus, the type of fatty acid (unsaturated or saturated) in the phospholipid did not influence the assimilation. As the amount of Tween 80 in the growth media increased beyond 0.05%, cholesterol uptake decreased, and the amount of growth remained the same. The higher concentrations of Tween 80 may have adversely affected the permeability of the cells

  13. Metabolic engineering of Corynebacterium glutamicum strain ATCC13032 to produce L-methionine.

    Science.gov (United States)

    Qin, Tianyu; Hu, Xiaoqing; Hu, Jinyu; Wang, Xiaoyuan

    2015-01-01

    L-Methionine-producing strain QW102/pJYW-4-hom(m) -lysC(m) -brnFE was developed from Corynebacterium glutamicum strain ATCC13032, using metabolic engineering strategies. These strategies involved (i) deletion of the gene thrB encoding homoserine kinase to increase the precursor supply, (ii) deletion of the gene mcbR encoding the regulator McbR to release the transcriptional repression to various genes in the l-methionine biosynthetic pathway, (iii) overexpression of the gene lysC(m) encoding feedback-resistant aspartate kinase and the gene hom(m) encoding feedback-resistant homoserine dehydrogenase to further increase the precursor supply, and (iv) overexpression of the gene cluster brnF and brnE encoding the export protein complex BrnFE to increase extracellular l-methionine concentration. QW102/pJYW-4-hom(m) -lysC(m) -brnFE produced 42.2 mM (6.3 g/L) l-methionine after 64-H fed-batch fermentation. These results suggest that l-methionine-producing strains can be developed from wild-type C. glutamicum strains by rationally metabolic engineering.

  14. Pivotal role of anthranilate dioxygenase genes in the adaptation of Burkholderia multivorans ATCC 17616 in soil.

    Science.gov (United States)

    Nishiyama, Eri; Ohtsubo, Yoshiyuki; Yamamoto, Yasuhiro; Nagata, Yuji; Tsuda, Masataka

    2012-05-01

    In our recent screen for soil-induced genes, the expression of andA operon (andAcAdAbAa) for anthranilate catabolism in Burkholderia multivorans ATCC 17616 was found to increase dramatically in a soil sample (Nishiyama et al., Environ Microbiol 12: 2539, 2010). The operon was preceded by andR encoding a putative transcriptional regulator for the andA operon. In this study, the andA promoter was induced by tryptophan and anthranilate in an andR-dependent manner. The andA promoter in a deletion mutant lacking tryptophan dioxygenase (one of enzymes for the catabolism of tryptophan to anthranilate) did not respond to tryptophan, indicating that not tryptophan but anthranilate is the effector of AndR. Although both anthranilate and tryptophan were under the detection levels in the soil sample, andA promoter showed higher activity in the soil sample than in a laboratory medium. Such induction required andR and was moderately dependent on the ferric uptake regulator (Fur). The proliferation ability of andAc mutant in the sterile soil was low compared with the co-incubated wild-type cells. These findings suggested that in the soil environment, anthranilate dioxygenase genes are induced by AndR and Fur, and play a pivotal role in the proliferation in the soil environment. PMID:22360670

  15. Ca2+/calmodulin dependent protein kinase from Mycobacterium smegmatis ATCC 607.

    Science.gov (United States)

    Sharma, S; Giri, S; Khuller, G K

    1998-06-01

    A soluble Ca2+/calmodulin dependent protein kinase has been partially purified (approximately 400 fold) from Mycobacterium smegmatis ATCC 607 using several purification steps like ammonium sulphate precipitation (30-60%), Sepharose CL-6B gel filtration, DEAE-cellulose and finally calmodulin-agarose affinity chromatography. On SDS-PAGE, this enzyme preparation showed a major protein band of molecular mass 35 kD and its activity was dependent on calcium, calmodulin and ATP when measured under saturating histone IIs (exogenous substrate) concentration. Phosphorylation of histone IIs was inhibited by W-7 (calmodulin inhibitor) and KN-62 (CaM-kinase inhibitor) with IC50 of 1.5 and 0.25 microm respectively, but was not affected by inhibitors of PKA (Sigma P5015) and PKC (H-7). All these results confirm that purified enzyme is Ca2+/calmodulin dependent protein kinase of M. smegmatis. The protein kinase of M. smegmatis demonstrated a narrow substrate specificity for both exogenous as well as endogenous substrates. These results suggest that purified CaM-kinase must be involved in regulating specific function(s) in this organism. PMID:9655195

  16. Uso do açafrão (Curcuma longa L. na redução da Escherichia coli (ATCC 25922 e Enterobacter aerogenes (ATCC 13048 em ricota The use of turmeric in the reduction of Escherichia coli (ATCC 25922 and Enterobacter aerogenes (ATCC 13048 in ricotta

    Directory of Open Access Journals (Sweden)

    Sandra Ribeiro Maia

    2004-04-01

    Full Text Available Considerando o envolvimento de queijos como veículo de microrganismos patogênicos, foi avaliada a eficiência do extrato alcoólico de cúrcuma adicionado à ricota, na redução de Escherichia coli e Enterobacter aerogenes. Foram fabricados três lotes de ricota cremosa e inoculados com 104 UFC/mL de Escherichia coli (ATCC 25922 e 105 UFC/mL de Enterobacter aerogenes (ATCC 13048. Às ricotas, foram adicionados 0,4% de NaCl e extrato alcoólico de Curcuma longa L., em concentrações que variaram de 0,0% a 2,0%. As ricotas foram avaliadas físico-química e microbiologicamente em 0, 1, 7, 14 e 21 dias de armazenamento refrigerado. O percentual de umidade das ricotas foi, em média, de 73%. O pH médio observado foi de 5,4 e o percentual de gordura de 3%. Pelos resultados, evidenciou-se, após 21 dias, uma redução do número de Escherichia coli de aproximadamente dois ciclos logaritmicos nos tratamentos utilizados de 0,5%, 1,0%, 1,5% e 2,0% de cúrcuma. Já para Enterobacter aerogenes, a redução foi menor, de aproximadamente um ciclo logaritmico, de 105 UFC/mL para 104 UFC/mL, também nos tratamentos utilizados de 0,5%, 1,0%, 1,5% e 2,0% de cúrcuma. Apesar de os resultados evidenciarem uma redução do número de células viáveis dos microrganismos avaliados, a cúrcuma não deverá ser o único meio preservativo, considerando uma contaminação inicial de 104 UFC/mL de Escherichia coli e 105 UFC/mL de Enterobacter aerogenes, pois não atenderia à legislação vigente quanto aos requisitos microbiológicos para queijos.Considering the cheese involvement as a vehicle of pathogenic microorganisms it was evaluated the eficciency of the ethanolic turmeric extract added to ricotta, in the reduction of Escherichia coli and Enterobacter aerogenes. Three lots of creamy ricotta were manufacturated and inoculated with 104 UFC/mL of Escherichia coli (ATCC 25922 and 105 UFC/mL of Enterobacter aerogenes (ATCC 13048. It was added 0,4% of NaCl and

  17. Effect of Low Shear Modeled Microgravity (LSMMG) on the Probiotic Lactobacillus Acidophilus ATCC 4356

    Science.gov (United States)

    Stahl, S.; Voorhies, A.; Lorenzi, H.; Castro-Wallace, S.; Douglas, G.

    2016-01-01

    The introduction of generally recognized as safe (GRAS) probiotic microbes into the spaceflight food system has the potential for use as a safe, non-invasive, daily countermeasure to crew microbiome and immune dysregulation. However, the microgravity effects on the stress tolerances and genetic expression of probiotic bacteria must be determined to confirm translation of strain benefits and to identify potential for optimization of growth, survival, and strain selection for spaceflight. The work presented here demonstrates the translation of characteristics of a GRAS probiotic bacteria to a microgravity analog environment. Lactobacillus acidophilus ATCC 4356 was grown in the low shear modeled microgravity (LSMMG) orientation and the control orientation in the rotating wall vessel (RWV) to determine the effect of LSMMG on the growth, survival through stress challenge, and gene expression of the strain. No differences were observed between the LSMMG and control grown L. acidophilus, suggesting that the strain will behave similarly in spaceflight and may be expected to confer Earth-based benefits.

  18. Metabolic engineering of Corynebacterium glutamicum ATCC13869 for L-valine production.

    Science.gov (United States)

    Chen, Cheng; Li, Yanyan; Hu, Jinyu; Dong, Xunyan; Wang, Xiaoyuan

    2015-05-01

    In this study, an L-valine-producing strain was developed from Corynebacterium glutamicum ATCC13869 through deletion of the three genes aceE, alaT and ilvA combined with the overexpression of six genes ilvB, ilvN, ilvC, lrp1, brnF and brnE. Overexpression of lrp1 alone increased L-valine production by 16-fold. Deletion of the aceE, alaT and ilvA increased L-valine production by 44-fold. Overexpression of the six genes ilvB, ilvN, ilvC, lrp1, brnE and brnF in the triple deletion mutant WCC003 further increased L-valine production. The strain WCC003/pJYW-4-ilvBNC1-lrp1-brnFE produced 243mM L-valine in flask cultivation and 437mM (51g/L) L-valine in fed-batch fermentation and lacked detectable amino-acid byproduct such as l-alanine and l-isoleucine that are usually found in the fermentation of L-valine-producing C. glutamicum.

  19. Heterologous expression and localization of gentisate transporter Ncg12922 from Corynebacterium glutamicum ATCC 13032

    International Nuclear Information System (INIS)

    Ralstonia sp. strain U2 metabolizes naphthalene via gentisate (2,5-dihydroxybenzoate) to central metabolites, but it was found unable to utilize gentisate as growth substrate. A putative gentisate transporter encoded by ncg12922 from Corynebacterium glutamicum ATCC 13032 was functionally expressed in Ralstonia sp. strain U2, converting strain U2 to a gentisate utilizer. After ncg12922 was inserted into plasmid pGFPe with green fluorescence protein gene gfp, the expressed fusion protein Ncg12922-GFP could be visualized in the periphery of Escherichia coli cells under confocal microscope, consistent with a cytoplasmic membrane location. In contrast, GFP was ubiquitous in the cytoplasm of E. coli cells carrying pGFPe only. Gentisate 1,2-dioxygenase activity was present in the cell extract from strain U2 induced with gentisate but at a much lower level (one-fifth) than that obtained with salicylate. However, it exhibited a similar level in strain U2 containing Ncg12922 induced either by salicylate or gentisate

  20. Mutation of aspartic acid residues in the fructosyltransferase of Streptococcus salivarius ATCC 25975.

    Science.gov (United States)

    Song, D D; Jacques, N A

    1999-01-01

    The site-directed mutated fructosyltransferases (Ftfs) of Streptococcus salivarius ATCC 25975, D312E, D312S, D312N and D312K were all active at 37 degrees C, indicating that Asp-312 present in the 'sucrose box' was not the nucleophilic Asp residue responsible for the formation of a covalent fructosyl-enzyme intermediate required for enzyme activity. Analysis of the kinetic constants of the purified mutated forms of the enzyme showed that Asp-312 was most likely an essential amino acid involved in determining acceptor recognition and/or stabilizing a beta-turn in the protein. In contrast, when the Asp-397 of the Ftf present in the conserved triplet RDP motif of all 60 bacterial and plant family-32 glycosylhydrolases was mutated to a Ser residue, both sucrose hydrolysis and polymerization ceased. Tryptophan emission spectra confirmed that this mutation did not alter protein structure. Comparison of published data from other site-directed mutated enzymes implicated the Asp residue in the RDP motif as the one that may form a transient covalent fructosyl intermediate during the catalysis of sucrose by the Ftf of S. salivarius. PMID:10548559

  1. Evaluation of Cyanothece sp. ATCC 51142 as a candidate for inclusion in a CELSS

    Science.gov (United States)

    Schneegurt, M. A.; Arieli, B.; Nielsen, S. S.; Trumbo, P. R.; Sherman, L. A.; Mitchell, C. A. (Principal Investigator)

    1996-01-01

    Controlled ecological life support systems (CELSS) have been proposed to make long-duration manned space flights more cost-effective. Higher plants will presumably provide food and a breathable atmosphere for the crew. It has been suggested that imbalances between the CO2/O2 gas exchange ratios of the heterotrophic and autotrophic components of the system will inevitably lead to an unstable system, and the loss of O2 from the atmosphere. Ratio imbalances may be corrected by including a second autotroph with an appropriate CO2/O2 gas exchange ratio. Cyanothece sp. ATCC 51142 is a large unicellular N2-fixing cyanobacterium, exhibiting high growth rates under diverse physiological conditions. A rat-feeding study showed the biomass to be edible. Furthermore, it may have a CO2/O2 gas exchange ratio that theoretically can compensate for ratio imbalances. It is suggested that Cyanothece spp. could fulfill several roles in a CELSS: supplementing atmosphere recycling, generating fixed N from the air, providing a balanced protein supplement, and protecting a CELSS in case of catastrophic crop failure.

  2. Effect of Environmental Parameters on hydrogen Production using Clostridium Saccharoperbutylacetonicum N1-4(ATCC 13564

    Directory of Open Access Journals (Sweden)

    Walid M. Alalayah

    2009-01-01

    Full Text Available Problem statement: Hydrogen gas production by Clostridium can be improved by several ways through media formulation, or suitable environment condition. This study was carried out to investigate the environmental factors effects on hydrogen production using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564. Approach: The environmental factor studied includes initial substrate concentration, initial medium pH, temperature, sparging nitrogen and addition of Fe2+. Results: The result showed that the best yield of hydrogen produced (YP/S was 3.10 moL (moL glucose-1 when an initial glucose concentration was 10 g L-1, initial pH 6.0±0.2 at temperature 37°C. The volume of hydrogen produced was decreased when higher initial glucose concentration was applied. The yield of hydrogen increased when Fe2+ added to medium at concentration of 25 mg L-1. The yield and growth were further increased by sparging with nitrogen gas. Conclusion: It was observed that the best condition for highest hydrogen yield when initial pH 6.0±0.2 at 37°C and enhanced by adding ferrous sulfate in anaerobic process.

  3. Effect of fermentation conditions on the production of hyaluronic acid by Streptococcus zooepidemicus ATCC 39920

    Directory of Open Access Journals (Sweden)

    Nicole Caldas Pan

    2015-10-01

    Full Text Available The production of hyaluronic acid by Streptococcus zooepidemicus ATCC 39920 with varying rates of pH (6.0, 7.0, 8.0, temperature (34; 37; 40°C, agitation (100, 150, 200 rpm, glucose (10, 20, 30 g L-1 and yeast extract concentration (10, 20, 30 g L-1 was evaluated by statistical approaches. The best conditions for the production of hyaluronic acid was pH 8.0, 37°C and 100 rpm in a medium containing 30 g L-1 glucose and yeast extract, for a production of 0.787 g L-1. Temperature, pH and yeast extract were significant variables (p < 0.05. Yeast extract and pH had a positive effect on the production of the polymer. Lactate, formate and acetate synthesis were also analyzed. Current assay showed the feasibility of statistical tools to optimize the physical and nutritional parameters for the production of hyaluronic acid and the improvement of the fermentation process.

  4. Staphylococcus saprophyticus ATCC 15305 is internalized into human urinary bladder carcinoma cell line 5637.

    Science.gov (United States)

    Szabados, Florian; Kleine, Britta; Anders, Agnes; Kaase, Martin; Sakinç, Türkân; Schmitz, Inge; Gatermann, Sören

    2008-08-01

    Invasion of bacteria into nonphagocytic host cells is an important pathogenicity factor for escaping the host defence system. Gram-positive organisms, for example Staphylococcus aureus and Listeria monocytogenes, are invasive in nonphagocytic cells, and this mechanism is discussed as an important part of the infection process. Uropathogenic Escherichia coli and Staphylococcus saprophyticus can cause acute and recurrent urinary tract infections as well as bloodstream infections. Staphylococcus saprophyticus shows strong adhesion to human urinary bladder carcinoma and Hep2 cells and expresses the 'Microbial Surface Components Recognizing Adhesive Matrix molecule' (MSCRAMM)-protein SdrI with collagen-binding activity. MSCRAMMs are responsible for adhesion and collagen binding in S. aureus and are discussed as an important pathogenicity factor for invasion. To investigate internalization in S. aureus, several fluorescence activated cell sorting (FACS) assays have been described recently. We used a previously described FACS assay, with slight modifications, in addition to an antibiotic protection assay and transmission electron microscopy to show that S. saprophyticus ATCC 15305 and the wild-type strain 7108 were internalized into the human urinary bladder carcinoma cell line 5637. The discovery of the internalization of S. saprophyticus may be an important step for understanding the pathogenicity of recurrent infections caused by this organism.

  5. Clavulanic acid production by the MMS 150 mutant obtained from wild type Streptomyces clavuligerus ATCC 27064

    Directory of Open Access Journals (Sweden)

    Eliton da Silva Vasconcelos

    2013-12-01

    Full Text Available Clavulanic acid (CA is a powerful inhibitor of the beta-lactamases, enzymes produced by bacteria resistants to penicillin and cefalosporin. This molecule is produced industrially by strains of Streptomyces clavuligerus in complex media which carbon and nitrogen resources are supplied by inexpensive compounds still providing high productivity. The genetic production improvement using physical and chemical mutagenic agents is an important strategy in programs of industrial production development of bioactive metabolites. However, parental strains are susceptible to loss of their original productivity due genetic instability phenomenona. In this work, some S. clavuligerus mutant strains obtained by treatment with UV light and with MMS are compared with the wild type (Streptomyces clavuligerus ATCC 27064. The results indicated that the random mutations originated some strains with different phenotypes, most divergent demonstrated by the mutants strains named AC116, MMS 150 and MMS 54, that exhibited lack of pigmentation in their mature spores. Also, the strain MMS 150 presented a larger production of CA when cultivated in semi-synthetics media. Using other media, the wild type strain obtained a larger CA production. Besides, using the modifed complex media the MMS 150 strain showed changes in its lipolitic activity and a larger production of CA. The studies also allowed finding the best conditions for a lipase activity exhibited by wild type S. clavuligerus and the MMS150 mutant.

  6. Clavulanic acid production by the MMS 150 mutant obtained from wild type Streptomyces clavuligerus ATCC 27064.

    Science.gov (United States)

    da Silva Vasconcelos, Eliton; de Lima, Vanderlei Aparecido; Goto, Leandro Seiji; Cruz-Hernández, Isara Lourdes; Hokka, Carlos Osamu

    2013-12-01

    Clavulanic acid (CA) is a powerful inhibitor of the beta-lactamases, enzymes produced by bacteria resistants to penicillin and cefalosporin. This molecule is produced industrially by strains of Streptomyces clavuligerus in complex media which carbon and nitrogen resources are supplied by inexpensive compounds still providing high productivity. The genetic production improvement using physical and chemical mutagenic agents is an important strategy in programs of industrial production development of bioactive metabolites. However, parental strains are susceptible to loss of their original productivity due genetic instability phenomenona. In this work, some S. clavuligerus mutant strains obtained by treatment with UV light and with MMS are compared with the wild type (Streptomyces clavuligerus ATCC 27064). The results indicated that the random mutations originated some strains with different phenotypes, most divergent demonstrated by the mutants strains named AC116, MMS 150 and MMS 54, that exhibited lack of pigmentation in their mature spores. Also, the strain MMS 150 presented a larger production of CA when cultivated in semi-synthetics media. Using other media, the wild type strain obtained a larger CA production. Besides, using the modifed complex media the MMS 150 strain showed changes in its lipolitic activity and a larger production of CA. The studies also allowed finding the best conditions for a lipase activity exhibited by wild type S. clavuligerus and the MMS150 mutant.

  7. Isolation and characterisation of dipeptidyl peptidase IV from Prevotella loescheii ATCC 15930.

    Science.gov (United States)

    Koreeda, Y; Hayakawa, M; Ikemi, T; Abiko, Y

    2001-08-01

    A proline-specific dipeptidyl aminopeptidase, dipeptidyl peptidase IV (EC 3.4.14.5), was purified from a cell sonicate soluble fraction of Prevotella loescheii ATCC 15930 by sequential column chromatography. The molecular mass of the native enzyme was estimated as 160 kDa by high-pressure liquid gel filtration column chromatography and unheated sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The subunit molecular mass was 80 kDa when the enzyme was heated to 100 degrees C in the presence of 2-mercaptoethanol before SDS-PAGE, suggesting that the native enzyme consists of two identical subunits and is folded in 2% SDS. The optimum pH, with glycyl-prolyl-4-methyl-coumaryl-7-amide as the substrate, was 8.0; the isoelectric point was 5.2. Purified enzyme showed a strong preference for dipeptide substrates containing proline and, less efficiently, alanine in the P1 position. The enzyme was markedly inhibited by Cd(2+), Zn(2+), Hg(2+), Co(2+), and serine proteinase inhibitor di-isopropylfluorophosphate. PMID:11389867

  8. Optimization of probiotic lactobacillus casei ATCC 334 production using date powder as carbon source

    Directory of Open Access Journals (Sweden)

    Shahravy A.

    2012-01-01

    Full Text Available This study was conducted to optimize culture conditions for economic production of a probiotic bacterium, Lactobacillus casei ATCC 334, in which palm date powder was applied for the first time as a low-cost main carbon source. The effect of eleven factors on bacterial growth was investigated using the Taguchi experimental design, and three factors including palm date powder, tryptone and agitation rate were found to be the most significant parameters. The optimum conditions including date powder concentration, 38 g/L; tryptone concentration, 30 g/L; and an agitation rate of 320 rpm were determined by response surface methodology of Box-Behnken. A third-order polynomial model was suggested to predict the design space following which the predicted values were validated experimentally. The maximum log value of the viable cells in the optimized alternative medium was 9.97 at 24 h of incubation which was comparable to that obtained in the complex and expensive MRS medium (10.06.

  9. Proteome data to explore the impact of pBClin15 on Bacillus cereus ATCC 14579.

    Science.gov (United States)

    Madeira, Jean-Paul; Alpha-Bazin, Béatrice; Armengaud, Jean; Omer, Hélène; Duport, Catherine

    2016-09-01

    This data article reports changes in the cellular and exoproteome of B. cereus cured from pBClin15.Time-course changes of proteins were assessed by high-throughput nanoLC-MS/MS. We report all the peptides and proteins identified and quantified in B. cereus with and without pBClin15. Proteins were classified into functional groups using the information available in the KEGG classification and we reported their abundance in term of normalized spectral abundance factor. The repertoire of experimentally confirmed proteins of B. cereus presented here is the largest ever reported, and provides new insights into the interplay between pBClin15 and its host B. cereus ATCC 14579. The data reported here is related to a published shotgun proteomics analysis regarding the role of pBClin15, "Deciphering the interactions between the Bacillus cereus linear plasmid, pBClin15, and its host by high-throughput comparative proteomics" Madeira et al. [1]. All the associated mass spectrometry data have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository (http://www.ebi.ac.uk/pride/), with the dataset identifier PRIDE: PXD001568, PRIDE: PXD002788 and PRIDE: PXD002789. PMID:27547804

  10. Investigation of the Amycolatopsis sp. Strain ATCC 39116 Vanillin Dehydrogenase and Its Impact on the Biotechnical Production of Vanillin

    OpenAIRE

    Fleige, Christian; Hansen, Gunda; Kroll, Jens; Steinbüchel, Alexander

    2013-01-01

    The actinomycete Amycolatopsis sp. strain ATCC 39116 is capable of synthesizing large amounts of vanillin from ferulic acid, which is a natural cell wall component of higher plants. The desired intermediate vanillin is subject to undesired catabolism caused by the metabolic activity of a hitherto unknown vanillin dehydrogenase (VDHATCC 39116). In order to prevent the oxidation of vanillin to vanillic acid and thereby to obtain higher yields and concentrations of vanillin, the responsible vani...

  11. Genomic and Genetic Characterization of the Bile Stress Response of Probiotic Lactobacillus reuteri ATCC 55730▿ †

    OpenAIRE

    Whitehead, Kristi; Versalovic, James; Roos, Stefan; Britton, Robert A.

    2008-01-01

    Probiotic bacteria encounter various stresses after ingestion by the host, including exposure to the low pH in the stomach and bile in the small intestine. The probiotic microorganism Lactobacillus reuteri ATCC 55730 has previously been shown to survive in the human small intestine. To address how L. reuteri can resist bile stress, we performed microarray experiments to determine gene expression changes that occur when the organism is exposed to physiological concentrations of bile. A wide va...

  12. Microbial Corrosion of API 5L X-70 Carbon Steel by ATCC 7757 and Consortium of Sulfate-Reducing Bacteria

    OpenAIRE

    2014-01-01

    Various cases of accidents involving microbiology influenced corrosion (MIC) were reported by the oil and gas industry. Sulfate reducing bacteria (SRB) have always been linked to MIC mechanisms as one of the major causes of localized corrosion problems. In this study, SRB colonies were isolated from the soil in suspected areas near the natural gas transmission pipeline in Malaysia. The effects of ATCC 7757 and consortium of isolated SRB upon corrosion on API 5L X-70 carbon steel coupon were i...

  13. Molecular cloning and characterization of the aklavinone 11-hydroxylase gene of Streptomyces peucetius subsp. caesius ATCC 27952.

    OpenAIRE

    Hong, Y S; Hwang, C K; Hong, S. K.; Kim, Y.H.(Center for Underground Physics, Institute for Basic Science (IBS), Daejon, 305-811, Korea); Lee, J. J.

    1994-01-01

    The gene encoding aklavinone 11-hydroxylase of Streptomyces peucetius subsp. caesius ATCC 27952 was cloned and sequenced. The deduced amino acid sequence of the gene contains at least two common motifs of well-conserved amino acid sequences of several flavin-type bacterial hydroxylases. The hydroxylase gene is apparently transcribed from a single transcriptional start point. The phenotype of a dnrF mutant generated by gene disruption supports the idea that the dnrF gene encodes aklavinone 11-...

  14. In vitro and in vivo activities of ticarcillin-loaded nanoliposomes with different surface charges against Pseudomonas aeruginosa (ATCC 29248

    Directory of Open Access Journals (Sweden)

    Gharib Amir

    2012-10-01

    Full Text Available Abstract Background Pseudomonas aeruginosa exhibits multiple antibiotic resistance mechanisms. Different studies have shown that entrapment of antibiotics into liposomes could increase their anti-Pseudomonas activity. The objectives of this study were to prepare ticarcillin loaded-nanoliposomes with variable surface charges and evaluate their in vitro and in vivo efficacies against Pseudomonas aeruginosa (ATCC 29248. Methods Ticarcillin-loaded nanoliposomes with positive, negative and neutral surface charges were prepared by extrusion method. Ticarcillin encapsulation efficacies for different formulations were measured by HPLC method. Minimum inhibitory concentration (MIC of ticarcillin nanoliposomal forms against strain ATCC 29248 were determined by broth dilution method. The killing rate of Pseudomonas aeruginosa was exposed to various concentrations of ticarcillin in free and nanoliposomal forms were analyzed. Ultimately, in vivo therapeutic efficacy of nanoliposomes in burned mice skin infected with strain ATCC 29248 was investigated. Results The encapsulation efficacies for ticarcillin-loaded cationic nanoliposomes were significantly higher (76% ± 0.17 than those of neutral (55% ± 0.14 and anionic (43% ± 0.14 nanoliposomes. The MIC of free, cationic, neutral and anionic nanoliposomal forms of ticarcillin against ATCC 29248 were to 24, 3, 6 and 48 mg/L, respectively. The killing rates of ticarcillin-loaded cationic nanoliposomes were higher than those of free and other drug formulations. Treatment by ticarcillin-loaded nanoliposomes with positive, neutral and negative surface charges resulted in almost 100, 60 and 20% survival rates, respectively. Conclusion Our data suggested that cationic ticarcillin-loaded nanoliposomes because of high effectiveness would be a good choice to treatment of Pseudomonas aeruginosa infections.

  15. Selection of the Strain Lactobacillus acidophilus ATCC 43121 and Its Application to Brewers' Spent Grain Conversion into Lactic Acid.

    Science.gov (United States)

    Liguori, Rossana; Soccol, Carlos Ricardo; Vandenberghe, Luciana Porto de Souza; Woiciechowski, Adenise Lorenci; Ionata, Elena; Marcolongo, Loredana; Faraco, Vincenza

    2015-01-01

    Six Lactobacillus strains were analyzed to select a bacterium for conversion of brewers' spent grain (BSG) into lactic acid. Among the investigated strains, L. acidophilus ATCC 43121 showed the highest yield of lactic acid production (16.1 g/L after 48 hours) when grown in a synthetic medium. It was then analyzed for its ability to grow on the hydrolysates obtained from BSG after acid-alkaline (AAT) or aqueous ammonia soaking (AAS) pretreatment. The lactic acid production by L. acidophilus ATCC 43121 through fermentation of the hydrolysate from AAS treated BSG was 96% higher than that from the AAT treated one, although similar yields of lactic acid per consumed glucose were achieved due to a higher (46%) glucose consumption by L. acidophilus ATCC 43121 in the AAS BSG hydrolysate. It is worth noting that adding yeast extract to the BSG hydrolysates increased both the yield of lactic acid per substrate consumed and the volumetric productivity. The best results were obtained by fermentation of AAS BSG hydrolysate supplemented by yeast extract, in which the strain produced 22.16 g/L of lactic acid (yield of 0.61 g/g), 27% higher than the value (17.49 g/L) obtained in the absence of a nitrogen source.

  16. Selection of the Strain Lactobacillus acidophilus ATCC 43121 and Its Application to Brewers’ Spent Grain Conversion into Lactic Acid

    Directory of Open Access Journals (Sweden)

    Rossana Liguori

    2015-01-01

    Full Text Available Six Lactobacillus strains were analyzed to select a bacterium for conversion of brewers’ spent grain (BSG into lactic acid. Among the investigated strains, L. acidophilus ATCC 43121 showed the highest yield of lactic acid production (16.1 g/L after 48 hours when grown in a synthetic medium. It was then analyzed for its ability to grow on the hydrolysates obtained from BSG after acid-alkaline (AAT or aqueous ammonia soaking (AAS pretreatment. The lactic acid production by L. acidophilus ATCC 43121 through fermentation of the hydrolysate from AAS treated BSG was 96% higher than that from the AAT treated one, although similar yields of lactic acid per consumed glucose were achieved due to a higher (46% glucose consumption by L. acidophilus ATCC 43121 in the AAS BSG hydrolysate. It is worth noting that adding yeast extract to the BSG hydrolysates increased both the yield of lactic acid per substrate consumed and the volumetric productivity. The best results were obtained by fermentation of AAS BSG hydrolysate supplemented by yeast extract, in which the strain produced 22.16 g/L of lactic acid (yield of 0.61 g/g, 27% higher than the value (17.49 g/L obtained in the absence of a nitrogen source.

  17. Microbial Corrosion of API 5L X-70 Carbon Steel by ATCC 7757 and Consortium of Sulfate-Reducing Bacteria

    Directory of Open Access Journals (Sweden)

    Arman Abdullah

    2014-01-01

    Full Text Available Various cases of accidents involving microbiology influenced corrosion (MIC were reported by the oil and gas industry. Sulfate reducing bacteria (SRB have always been linked to MIC mechanisms as one of the major causes of localized corrosion problems. In this study, SRB colonies were isolated from the soil in suspected areas near the natural gas transmission pipeline in Malaysia. The effects of ATCC 7757 and consortium of isolated SRB upon corrosion on API 5L X-70 carbon steel coupon were investigated using a weight loss method, an open circuit potential method (OCP, and a potentiodynamic polarization curves method in anaerobic conditions. Scanning electron microscopy (SEM and energy dispersive X-ray spectroscopy (EDS were then used to determine the corrosion morphology in verifying the SRB activity and corrosion products formation. Results from the study show that the corrosion rate (CR of weight loss method for the isolated SRB is recorded as 0.2017 mm/yr compared to 0.2530 mm/yr for ATCC 7757. The Tafel plot recorded the corrosion rate of 0.3290 mm/yr for Sg. Ular SRB and 0.2500 mm/yr for Desulfovibrio vulgaris. The results showed that the consortia of isolated SRB were of comparable effects and features with the single ATCC 7757 strain.

  18. Lactobacillus acidophilus ATCC 4356 inhibits biofilm formation by C. albicans and attenuates the experimental candidiasis in Galleria mellonella.

    Science.gov (United States)

    Vilela, Simone F G; Barbosa, Júnia O; Rossoni, Rodnei D; Santos, Jéssica D; Prata, Marcia C A; Anbinder, Ana Lia; Jorge, Antonio O C; Junqueira, Juliana C

    2015-01-01

    Probiotic strains of Lactobacillus have been studied for their inhibitory effects on Candida albicans. However, few studies have investigated the effect of these strains on biofilm formation, filamentation and C. albicans infection. The objective of this study was to evaluate the influence of Lactobacillus acidophilus ATCC 4356 on C. albicans ATCC 18804 using in vitro and in vivo models. In vitro analysis evaluated the effects of L. acidophilus on the biofilm formation and on the capacity of C. albicans filamentation. For in vivo study, Galleria mellonella was used as an infection model to evaluate the effects of L. acidophilus on candidiasis by survival analysis, quantification of C. albicans CFU/mL, and histological analysis. The direct effects of L. acidophilus cells on C. albicans, as well as the indirect effects using only a Lactobacillus culture filtrate, were evaluated in both tests. The in vitro results showed that both L. acidophilus cells and filtrate were able to inhibit C. albicans biofilm formation and filamentation. In the in vivo study, injection of L. acidophilus into G. mellonella larvae infected with C. albicans increased the survival of these animals. Furthermore, the number of C. albicans CFU/mL recovered from the larval hemolymph was lower in the group inoculated with L. acidophilus compared to the control group. In conclusion, L. acidophilus ATCC 4356 inhibited in vitro biofilm formation by C. albicans and protected G. mellonella against experimental candidiasis in vivo.

  19. Antibacterial activity of antagonistic bacterium Bacillus subtilis DJM-51 against phytopathogenic Clavibacter michiganense subsp. michiganense ATCC 7429 in vitro.

    Science.gov (United States)

    Jung, W J; Mabood, F; Souleimanov, A; Whyte, L G; Niederberger, T D; Smith, D L

    2014-12-01

    To investigate antibacterial activity against the tomato pathogen Clavibacter michiganense subsp. michiganense ATCC 7429 (Cmm ATCC 7429), Bacillus subtilis DJM-51 was isolated from rhizosphere soil. For isolation of bacteria, samples were taken from rhizosphere soil. The isolate, DJA-51, had strong antagonistic ability against Tomato pathogen Cmm ATCC 7429 on nutrient-broth yeast extract agar (NBYA) as indicated by inhibition zones around colonies. On the basis of the nucleotide sequence of a conserved segment of the 16S rRNA gene, the bacterium has been identified as B. subtilis DJM-51. The growth of Cmm ATCC 7429 on NBYA plates was inhibited by culture broth of B. subtilis DJM-51 including cells, by the supernatant of culture broth of B. subtilis DJM-51, and by the liquid material resulting from butanol extract of bacterial cultures. The OD value in co-culture mixture was lower than the control throughout the entire incubation period. Antibiotics obtained from B. subtilis DJM-51 inhibited the growth of Tomato pathogen Cmm ATCC 7429. These results provide potentially information about the protection of tomato from pathogen Cmm ATCC 7429 under greenhouse conditions in Quebec. PMID:25457795

  20. Metabolic flux analysis of Cyanothece sp. ATCC 51142 under mixotrophic conditions.

    Science.gov (United States)

    Alagesan, Swathi; Gaudana, Sandeep B; Sinha, Avinash; Wangikar, Pramod P

    2013-11-01

    Cyanobacteria are a group of photosynthetic prokaryotes capable of utilizing solar energy to fix atmospheric carbon dioxide to biomass. Despite several "proof of principle" studies, low product yield is an impediment in commercialization of cyanobacteria-derived biofuels. Estimation of intracellular reaction rates by (13)C metabolic flux analysis ((13)C-MFA) would be a step toward enhancing biofuel yield via metabolic engineering. We report (13)C-MFA for Cyanothece sp. ATCC 51142, a unicellular nitrogen-fixing cyanobacterium, known for enhanced hydrogen yield under mixotrophic conditions. Rates of reactions in the central carbon metabolism under nitrogen-fixing and -non-fixing conditions were estimated by monitoring the competitive incorporation of (12)C and (13)C from unlabeled CO2 and uniformly labeled glycerol, respectively, into terminal metabolites such as amino acids. The observed labeling patterns suggest mixotrophic growth under both the conditions, with a larger fraction of unlabeled carbon in nitrate-sufficient cultures asserting a greater contribution of carbon fixation by photosynthesis and an anaplerotic pathway. Indeed, flux analysis complements the higher growth observed under nitrate-sufficient conditions. On the other hand, the flux through the oxidative pentose phosphate pathway and tricarboxylic acid cycle was greater in nitrate-deficient conditions, possibly to supply the precursors and reducing equivalents needed for nitrogen fixation. In addition, an enhanced flux through fructose-6-phosphate phosphoketolase possibly suggests the organism's preferred mode under nitrogen-fixing conditions. The (13)C-MFA results complement the reported predictions by flux balance analysis and provide quantitative insight into the organism's distinct metabolic features under nitrogen-fixing and -non-fixing conditions.

  1. DNA Sequence and Comparison of Virulence Plasmids from Rhodococcus equi ATCC 33701 and 103

    Science.gov (United States)

    Takai, Shinji; Hines, Stephen A.; Sekizaki, Tsutomu; Nicholson, Vivian M.; Alperin, Debra A.; Osaki, Makoto; Takamatsu, Daisuke; Nakamura, Mutsu; Suzuki, Kayo; Ogino, Nobuko; Kakuda, Tsutomu; Dan, Hanhong; Prescott, John F.

    2000-01-01

    The virulence plasmids of the equine virulent strains Rhodococcus equi ATCC 33701 and 103 were sequenced, and their genetic structure was analyzed. p33701 was 80,610 bp in length, and p103 was 1 bp shorter; their sequences were virtually identical. The plasmids contained 64 open reading frames (ORFs), 22 of which were homologous with genes of known function and 3 of which were homologous with putative genes of unknown function in other species. Putative functions were assigned to five ORFs based on protein family characteristics. The most striking feature of the virulence plasmids was the presence of a 27,536-bp pathogenicity island containing seven virulence-associated protein (vap) genes, including vapA. These vap genes have extensive homology to vapA, which encodes a thermoregulated and surface-expressed protein. The pathogenicity island contained a LysR family transcriptional regulator and a two-component response regulator upstream of six of the vap genes. The vap genes were present as a cluster of three (vapA, vapC, and vapD), as a pair (vapE and vapF), or individually (vapG; vapH). A region of extensive direct repeats of unknown function, possibly associated with thermoregulation, was present immediately upstream of the clustered and the paired genes but not the individual vap genes. There was extensive homology among the C-terminal halves of all vap genes but not generally among the N-terminal halves. The remainder of the plasmid consisted of a large region which appears to be associated with conjugation functions and a large region which appears to be associated with replication and partitioning functions. PMID:11083803

  2. Streptococcus salivarius ATCC 25975 possesses at least two genes coding for primer-independent glucosyltransferases.

    Science.gov (United States)

    Simpson, C L; Giffard, P M; Jacques, N A

    1995-01-01

    Fractionation of the culture medium showed that Streptococcus salivarius ATCC 25975 secreted a glucosyltransferase (Gtf) that was primer independent. On the basis of this observation, a gene library of S. salivarius chromosomal DNA cloned into lambda L47.1 was screened for a gene(s) coding for such an activity. As a result of this screening process, two new gtf genes, gtfL and gtfM, both of which coded for primer-independent Gtf activities, were isolated. GtfL produced an insoluble glucan that was refractory to digestion by the endo-(1-->6)-alpha-D-glucanase. of Chaetonium gracile, while GtfM produced a soluble glucan that was readily degraded by the glucanase. Comparison of the deduced amino acid sequences of gtfL and gtfM with 10 other available Gtf sequences allowed the relatedness of the conserved catalytic regions to be assessed. This analysis showed that the 12 enzymes did not form clusters based on their primer dependencies or on their product solubilities. Further analysis of the YG repeats in the C-terminal glucan-binding domains of GtfJ, GtfK, GtfL, and GtfM from S. salivarius showed that there was strong homology between a block of contiguous triplet YG repeats present in the four alleles. These blocks of YG repeats were coded for by a region of each gene that appeared to have arisen as a result of a recent duplication event(s). PMID:7822030

  3. Induction of secondary metabolism of Aspergillus terreus ATCC 20542 in the batch bioreactor cultures.

    Science.gov (United States)

    Boruta, Tomasz; Bizukojc, Marcin

    2016-04-01

    Cultivation of Aspergillus terreus ATCC 20542 in a stirred tank bioreactor was performed to induce the biosynthesis of secondary metabolites and provide the bioprocess-related insights into the metabolic capabilities of the investigated strain. The activation of biosynthetic routes was attempted by the diversification of process conditions and growth media. Several strategies were tested, including the addition of rapeseed oil or inulin, changing the concentration of nitrogen source, reduction of chlorine supply, cultivation under saline conditions, and using various aeration schemes. Fifteen secondary metabolites were identified in the course of the study by using ultra-high performance liquid chromatography coupled with mass spectrometry, namely mevinolinic acid, 4a,5-dihydromevinolinic acid, 3α-hydroxy-3,5-dihydromonacolin L acid, terrein, aspulvinone E, dihydroisoflavipucine, (+)-geodin, (+)-bisdechlorogeodin, (+)-erdin, asterric acid, butyrolactone I, desmethylsulochrin, questin, sulochrin, and demethylasterric acid. The study also presents the collection of mass spectra that can serve as a resource for future experiments. The growth in a salt-rich environment turned out to be strongly inhibitory for secondary metabolism and the formation of dense and compact pellets was observed. Generally, the addition of inulin, reducing the oxygen supply, and increasing the content of nitrogen source did not enhance the production of examined molecules. The most successful strategy involved the addition of rapeseed oil to the chlorine-deficient medium. Under these conditions, the highest levels of butyrolactone I, asterric acid, and mevinolinic acid were achieved and the presence of desmethylsulochrin and (+)-bisdechlorogeodin was detected in the broth. The constant and relatively high aeration rate in the idiophase was shown to be beneficial for terrein and (+)-geodin biosynthesis.

  4. Regulation of Three Nitrogenase Gene Clusters in the Cyanobacterium Anabaena variabilis ATCC 29413

    Directory of Open Access Journals (Sweden)

    Teresa Thiel

    2014-12-01

    Full Text Available The filamentous cyanobacterium Anabaena variabilis ATCC 29413 fixes nitrogen under aerobic conditions in specialized cells called heterocysts that form in response to an environmental deficiency in combined nitrogen. Nitrogen fixation is mediated by the enzyme nitrogenase, which is very sensitive to oxygen. Heterocysts are microxic cells that allow nitrogenase to function in a filament comprised primarily of vegetative cells that produce oxygen by photosynthesis. A. variabilis is unique among well-characterized cyanobacteria in that it has three nitrogenase gene clusters that encode different nitrogenases, which function under different environmental conditions. The nif1 genes encode a Mo-nitrogenase that functions only in heterocysts, even in filaments grown anaerobically. The nif2 genes encode a different Mo-nitrogenase that functions in vegetative cells, but only in filaments grown under anoxic conditions. An alternative V-nitrogenase is encoded by vnf genes that are expressed only in heterocysts in an environment that is deficient in Mo. Thus, these three nitrogenases are expressed differentially in response to environmental conditions. The entire nif1 gene cluster, comprising at least 15 genes, is primarily under the control of the promoter for the first gene, nifB1. Transcriptional control of many of the downstream nif1 genes occurs by a combination of weak promoters within the coding regions of some downstream genes and by RNA processing, which is associated with increased transcript stability. The vnf genes show a similar pattern of transcriptional and post-transcriptional control of expression suggesting that the complex pattern of regulation of the nif1 cluster is conserved in other cyanobacterial nitrogenase gene clusters.

  5. The Complete Genome Sequence of the Marine, Chemolithoautotrophic, Ammonia-Oxidizing Bacterium Nitrosococcus oceani ATCC19707

    Energy Technology Data Exchange (ETDEWEB)

    Klotz, M G; Arp, D J; Chain, P S; El-Sheikh, A F; Hauser, L J; Hommes, N G; Larimer, F W; Malfatti, S A; Norton, J M; Poret-Peterson, A T; Vergez, L M; Ward, B B

    2006-08-03

    The Gammaproteobacterium, Nitrosococcus oceani (ATCC 19707), is a Gram-negative obligate chemolithoautotroph capable of extracting energy and reducing power from the oxidation of ammonia to nitrite. Sequencing and annotation of the genome revealed a single circular chromosome (3,481,691 bp; 50.4% G+C) and a plasmid (40,420 bp) that contain 3052 and 41 candidate protein-encoding genes, respectively. The genes encoding proteins necessary for the function of known modes of lithotrophy and autotrophy were identified. In contrast to betaproteobacterial nitrifier genomes, the N. oceani genome contained two complete rrn operons. In contrast, only one copy of the genes needed to synthesize functional ammonia monooxygenase and hydroxylamine oxidoreductase, as well as the proteins that relay the extracted electrons to a terminal electron acceptor were identified. The N. oceani genome contained genes for 13 complete two-component systems. The genome also contained all the genes needed to reconstruct complete central pathways, the tricarboxylic acid cycle and the Embden-Meyerhof-Parnass and pentose phosphate pathways. The N. oceani genome contains the genes required to store and utilize energy from glycogen inclusion bodies and sucrose. Polyphosphate and pyrophosphate appear to be integrated in this bacterium's energy metabolism, stress tolerance and the ability to assimilate carbon via gluconeogenesis. One set of genes for type I RuBisCO was identified, while genes necessary for methanotrophy and for carboxysome formation were not identified. The N. oceani genome contains two copies each of the genes or operons necessary to assemble functional complexes I and IV as well as ATP synthase (one H{sup +}-dependent F{sub 0}F{sub 1}-type, one Na{sup +}-dependent V-type).

  6. 阿特拉津降解茵FM326(Arthrobacter sp.)的分离筛选、鉴定和生物学特性%Isolation, Screening and Identification of Atrazine-degrading Bacterium FM326(Arthrobacter sp.)

    Institute of Scientific and Technical Information of China (English)

    李明锐; 祖艳群; 陈建军; 湛方栋; 何永美; 李元

    2011-01-01

    采集除草剂阿特拉津污染的土壤,通过直接涂布法和富集驯化培养分离法,分别获得6株和5株能够降解阿特拉津的细菌.通过降解效率和降解动态试验,筛选到1株高效降解阿特拉津的菌株FM326,该菌株能以阿特拉津为唯一的碳源和氮源生长,培养96 h后对1000 mg· L-1阿特拉津降解效率达到97%.通过生理生化鉴定和16S rDNA序列分析,菌株FM326鉴定为节杆菌属(Arthrobacter sp.)细菌.该菌株表现出最适生长温度30~35℃,最适生长pH值5~9,好氧生长的生长特性.%The Iriazine herbicide atrazine(2-chloro-4-ethylamino-6-isopropyl-amino-s-triazine) is one of the most used pesticides. Atrazine is principally used for control of certain annual broadleaf and grass weeds, primarily in com but also in sorghum, sugarcane, and other crops. Although atrazine has relatively low persistency in soil, its remanets have been found in many higher concentrations even years after application due to widespread use. Microbial remediation is an effective and economic method to control the environment that has been polluted by atrazine. From herbicide contaminated soil, 5 atrazine degrading bacterial strains were isolated by enrichment culture, and 6 a- trazine-degrading bacterial strains were isolated by direct coating method. In the 11 strains, FM326 was screened out as the most highly efficient degradation bacteria by degradation test and could use atrazine as the sole carbon and nitrogen sources for growth. The degradation efficiency of strain FM326 was up to 97% within 96 hours while atrazine concentration was 1 000 mg-L"'. Strain FM326 was identified as Arthrobacter sp. According to physiological and biochemical tests and the phylogenic tree established by means of 16S rDNA sequencing. Moreover, strain FM326 was aerobic bacteria, its optimum temperature and pH value for growth was 30-35 t and 5~9, respectively. Strain of highly efficient degrading atrazine was found from soils

  7. Influence of single-walled carbon nanotubes on the growth and phenol biodegradation characteristics of Arthrobacter sp.W1%单壁碳纳米管对Arthrobacter sp.W1生长及苯酚降解功能的影响

    Institute of Scientific and Technical Information of China (English)

    李端行; 王经伟; 沈文丽; 张照婧; 厉舒祯; 李会杰; 刘紫嫣; 马桥; 曲媛媛

    2015-01-01

    单壁碳纳米管(SWCNTs)对大肠杆菌等模式菌株的生长抑制作用明显,为了解SWCNTs对具有环境功能的菌株的作用,以苯酚降解菌Arthrobacter sp.Wl为对象,考察不同浓度SWCNTs下菌株W1的生长曲线和苯酚降解曲线,并通过扫描电镜观察、细胞凋亡检测、DNA泄漏量分析和活性氧产量分析考察作用机制.结果表明,特定浓度范围的SWCNTs (0.5-5.0 mg/L)会加快W1的苯酚降解速率,且1.5-2.0 mg/L SWCNTs不抑制Wl的生长.SWCNTs在溶液中形成团簇并吸附Wl细胞,且对Wl产生以物理穿刺为主的毒性作用,但在特定浓度范围的SWCNTS条件下,SWCNTs-菌株-苯酚体系增加了菌体与苯酚的接触机会,从而对Wl生长和苯酚降解产生明显的促进作用.本研究结果可为进一步揭示SWCNTs的环境微生物效应提供理论依据.

  8. Optimization of Process Parameters of Producing Trehalose Synthase by Arthrobacter nicotinovorus Fermentation%响应面法优化食尼古丁节杆菌发酵产海藻糖合成酶的工艺

    Institute of Scientific and Technical Information of China (English)

    张良; 肖勇生; 包珍; 汤峰; 刘媛洁; 郭德斌

    2012-01-01

    [目的]应用响应面法优化食尼古丁节杆菌发酵产海藻糖合成酶的工艺.[方法]首先以培养温度、初始pH、摇床转速、接种量、发酵时间为因素进行单因素试验,在此基础上,应用 Box-Behnken 中心组合设计,以摇床转速、接种量和发酵时间为因素,以食尼古丁节杆菌发酵产海藻糖合成酶酶活的变化为响应值进行试验,运用SAS统计软件对试验数据进行分析,建立二次响应面回归模型,得出重要因素的最佳水平,从而确定最佳的发酵产酶条件.[结果]经响应面法优化获得食尼古丁节杆菌发酵产海藻糖合成酶的工艺参数为转速172.60 r/min、接种量13.96%、发酵时间55.97 h,在此优化条件下,海藻糖合成酶酶活力达到645.48 U/g.[结论]该研究可为海藻糖的生产提供科学依据.%[Objective] To optimize the process parameters of producing.trehalose synthase by Arthrobacter nicotinovorus fermentation. [ Method] The influence of fermentation temperature, initial pH, rotational speed, inoculation amount and fermentation time on trehalose synthase enzyme activity producing by Arthrobacter nicotinovorus fermentation were studied through single factor experiment, and approximately determined the a-mount of the various factors. Then the rotational speed, the inoculation amount and the fermentation time were selected as three factors, the center combination of Box-Behnken design experiments was used to study the change of trehalose synthase enzyme activity in the fermentation as response value. The experiment dates were analyzed by the SAS statistical software to set up quadratic response surface regression model for obtaining the the optimal level of the important factor, determining the optimum conditions of fermentation. [ Result ] The response surface method results showed that the optimum condition of producing trehalose synthase by Arthrobacter nicotinovorus fermentation was as follow; the rotational speed 172

  9. Crude fatty acid extracts of Streptomyces sps inhibits the biofilm forming Streptococcus pyogenes ATCC 19615

    Directory of Open Access Journals (Sweden)

    Rajalakshm Manickam

    2014-01-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Crude fatty acid extract of soil Streptomyces sps on the biofilm formation by Streptococcus pyogenes ATCC 19615 was investigated. Totally, 25 Streptomyces sps were isolated identified from the soil samples collected from Nilgiris hill station. All the isolates were subjected to hydrogen peroxide assay, fatty acid extraction and antibiofilm assay. The fatty acid extracts of S8, S9, and S15 inhibited S. pyogenes at MIC 10 µg/ml. The BIC was observed as 84.6% , 96.41%, 80.5% at 50 µg/ml concentration. Streptolysin S assay showed that the crude lipid extracts have the capability of inhibiting the Streptolysin S activity. There were changes in extracellular protein of the pathogen exposed to the S8, S9 and S15 crude fatty acid extracts (50 µg/ml at the range of 100-120 kDa which elucidates that the fatty acid extracts have a significant role in altering the extracellular protein which might be responsible for virulence of the pathogen. /* 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;}

  10. Significance of CO2 donor on the production of succinic acid by Actinobacillus succinogenes ATCC 55618

    Directory of Open Access Journals (Sweden)

    Zou Wei

    2011-10-01

    Full Text Available Abstract Background Succinic acid is a building-block chemical which could be used as the precursor of many industrial products. The dissolved CO2 concentration in the fermentation broth could strongly regulate the metabolic flux of carbon and the activity of phosphoenolpyruvate (PEP carboxykinase, which are the important committed steps for the biosynthesis of succinic acid by Actinobacillus succinogenes. Previous reports showed that succinic acid production could be promoted by regulating the supply of CO2 donor in the fermentation broth. Therefore, the effects of dissolved CO2 concentration and MgCO3 on the fermentation process should be investigated. In this article, we studied the impacts of gaseous CO2 partial pressure, dissolved CO2 concentration, and the addition amount of MgCO3 on succinic acid production by Actinobacillus succinogenes ATCC 55618. We also demonstrated that gaseous CO2 could be removed when MgCO3 was fully supplied. Results An effective CO2 quantitative mathematical model was developed to calculate the dissolved CO2 concentration in the fermentation broth. The highest succinic acid production of 61.92 g/L was obtained at 159.22 mM dissolved CO2 concentration, which was supplied by 40 g/L MgCO3 at the CO2 partial pressure of 101.33 kPa. When MgCO3 was used as the only CO2 donor, a maximal succinic acid production of 56.1 g/L was obtained, which was just decreased by 7.03% compared with that obtained under the supply of gaseous CO2 and MgCO3. Conclusions Besides the high dissolved CO2 concentration, the excessive addition of MgCO3 was beneficial to promote the succinic acid synthesis. This was the first report investigating the replaceable of gaseous CO2 in the fermentation of succinic acid. The results obtained in this study may be useful for reducing the cost of succinic acid fermentation process.

  11. Interaction of Wild Strains of Aspergilla with Aspergillus parasiticus ATCC15517 and Aflatoxin Production †

    Science.gov (United States)

    Martins, H. Marina; Almeida, Inês; Marques, Marta; Bernardo, Fernando

    2008-01-01

    Aflatoxins are secondary metabolites produced by some competent mould strains of Aspergillus flavus, A. parasiticus and A. nomius. These compounds have been extensively studied with regards to their toxicity for animals and humans; they are able to induce liver cancer and may cause a wide range of adverse effects in living organisms. Aflatoxins are found as natural contaminants of food and feed; the main line of the strategy to control them is based on the prevention of the mould growth in raw vegetable or during its storage and monitoring of each crop batch. Mould growth is conditioned by many ecological factors, including biotic ones. Hazard characterization models for aflatoxins in crops must take into consideration biotic interactions between moulds and their potential effects on growth development. The aim of this work is to study the effect of the biotic interaction of 14 different wild strains of Aspergilla (different species), with a competent strain (Aspergillus parasiticus ATCC 15517) using an in vitro production model. The laboratory model used was a natural matrix (humidified cracked corn), on which each wild strain challenged the aflatoxin production of a producer strain. Cultures were incubated at 28°C for 12 days and sampled at the 8th and 12th. Aflatoxin detection and quantification was performed by HPLC using a procedure with a MRPL = 1 μg/kg. Results of those interactive cultures revealed both synergic and antagonistic effects on aflatoxin biosynthesis. Productivity increases were particularly evident on the 8th day of incubation with wild strains of A. flavipes (+ 70.4 %), A. versicolor (+ 54.9 %) and A. flavus 3 (+ 62.6 %). Antagonistic effects were found with A. niger (− 69.5%), A. fumigatus (− 47.6 %) and A. terreus (− 47.6 %) on the 12th day. The increased effects were more evident on the 8th of incubation and the decreases were more patent on the 12th day. Results show that the development of Aspergilla strains concomitantly with

  12. The Biosynthesis of UDP-D-QuiNAc in Bacillus cereus ATCC 14579.

    Directory of Open Access Journals (Sweden)

    Soyoun Hwang

    Full Text Available N-acetylquinovosamine (2-acetamido-2,6-di-deoxy-D-glucose, QuiNAc is a relatively rare amino sugar residue found in glycans of few pathogenic gram-negative bacteria where it can play a role in infection. However, little is known about QuiNAc-related polysaccharides in gram-positive bacteria. In a routine screen for bacillus glycan grown at defined medium, it was surprising to identify a QuiNAc residue in polysaccharides isolated from this gram-positive bacterium. To gain insight into the biosynthesis of these glycans, we report the identification of an operon in Bacillus cereus ATCC 14579 that contains two genes encoding activities not previously described in gram-positive bacteria. One gene encodes a UDP-N-acetylglucosamine C4,6-dehydratase, (abbreviated Pdeg that converts UDP-GlcNAc to UDP-4-keto-4,6-D-deoxy-GlcNAc (UDP-2-acetamido-2,6-dideoxy-α-D-xylo-4-hexulose; and the second encodes a UDP-4-reductase (abbr. Preq that converts UDP-4-keto-4,6-D-deoxy-GlcNAc to UDP-N-acetyl-quinovosamine in the presence of NADPH. Biochemical studies established that the sequential Pdeg and Preq reaction product is UDP-D-QuiNAc as determined by mass spectrometry and one- and two-dimensional NMR experiments. Also, unambiguous evidence for the conversions of the dehydratase product, UDP-α-D-4-keto-4,6-deoxy-GlcNAc, to UDP-α-D-QuiNAc was obtained using real-time 1H-NMR spectroscopy and mass spectrometry. The two genes overlap by 4 nucleotides and similar operon organization and identical gene sequences were also identified in a few other Bacillus species suggesting they may have similar roles in the lifecycle of this class of bacteria important to human health. Our results provide new information about the ability of Bacilli to form UDP-QuiNAc and will provide insight to evaluate their role in the biology of Bacillus.

  13. Design and production of functionalized biopolyesters by Methylobacterium extorquens ATCC 55366: Toward new tissue engineering materials

    Science.gov (United States)

    Hoefer, Heinrich Friedrich Philipp Till Nikolaus

    Vascular networks are required to support the formation and function of three-dimensional tissues. Biodegradable scaffolds are being considered in order to promote vascularization where natural regeneration of lost or destroyed vascular networks fails. Particularly; composite materials are expected to fulfill the complex demands of a patient's body to support wound healing. Microbial biopolyesters are being regarded as such second and third generation biomaterials. Methylobacterium extorquens is one of several microorganisms that should be considered for the production of advanced polyhydroxyalkanoates (PHAs). M. extorquens displays a distinct advantage in that it is able to utilize methanol as an inexpensive substrate for growth and biopolyester production. The design of functionalized PHAs, which would be made of both saturated short-chain-length (scl, C ≤ 5) and unsaturated medium-chain-length (mcl, 6 ≤ C ≤ 14) monomeric units, aimed at combining desirable material properties of inert scl/mcl-PHAs with those of functionalized mcl-PHAs. By independently inserting the phaC1 or the phaC2 gene from Pseudomonas fluorescens GK13, recombinant M. extorquens strains were obtained which were capable of producing PHAs containing C-C double bonds. A fermentation process was developed to obtain gram quantities of biopolyesters employing the recombinant M. extorquens ATCC 55366 strain which harbored the phaC2 gene of P. fluorescens GK13, the better one of the two strains at incorporating unsaturated monomeric units. The PHAs produced were found in a blend of scl-PHAs and functionalized scl/mcl-PHAs (4 ≤ C ≤ 6), which were the products of the native and of the recombinant PHA synthase, respectively. Thermo-mechanical analysis confirmed that the functionalized scl/mcl-PHAs exhibited the desirable material properties expected. This project contributed to current research on polyhydroxyalkanoates at different levels. The terminal double bonds of the functionalized scl

  14. Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024

    DEFF Research Database (Denmark)

    Gallo, Giuseppe; Renzone, Giovanni; Palazzotto, Emilia;

    2016-01-01

    -dimensional difference in gel electrophoresis, mass spectrometry and gene ontology approaches. Microbispora ATCC-PTA-5024 cultivations in a complex medium were characterized by stages of biomass accumulation (A) followed by biomass yield decline (D). NAI-107 production started at 90 h (A stage), reached a maximum at 140...... differentiation of a TetR-like regulator, originally identified in this study, was confirmed by the construction of an over-expressing strain. Finally, the possible role of cellular response to membrane stability alterations and of multi-drug resistance ABC transporters as additional self-resistance mechanisms...

  15. EFFECT OF CULTURE MEDIUM ON BACTERIOCIN PRODUCTION BY LACTOBACILLUS RHAMNOSUS HN001 AND LACTOBACILLUS REUTERI ATCC 53608

    OpenAIRE

    Aguilar-Uscanga B. R.; Solís-Pacheco J. R.; Plascencia L.; Aguilar-Uscanga M. G.; García H. S.; Lacroix M.

    2013-01-01

    The aim of this study was to evaluate the effect of media on bacteriocin production by Lactobacillus rhamnosus HN001 and Lactobacillus reuteri ATCC 53608 using three different media: YPM, YPF and MRS supplemented with glucose and K2HPO4. The optimum temperature was 37°C and initial pH 6.5. Bacteriocin-like substances produced by tested bacteria in MRS medium supplemented with glucose and K2HPO4 exhibited a broad antimicrobial spectrum determined by well diffusion assay against indicator bacte...

  16. Growth of Cellulomonas sp. ATCC 21399 on different polysaccharides as sole carbon source induction of extracellular enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Poulsen, O.M.; Petersen, L.W.

    1988-11-01

    Growth and extracellular enzyme production of Cellulomonas sp. ATCC 21399 on carboxymethylcellulose (CMC), microcrystalline cellulose (Avicel), xylan, galactomannan and starch were compared. The bacteria grew poorly on CMC, whereas high cell densities were obtained on the other substrates. Growth on Avicel resulted in extrecellular enzyme activities against CMC, Avicel, xylan, galactomannan and amylose. By contrast, growth on xylan, galactomannan and starch induced only the enzymes neccessary for the degradation of the growth substrate. Extracellular proteinase activity could be measured during growth on all substrates but CMC, and the possibility of proteolytic inactivation of some of the unstable enzymes (i.e. Avicelase and amylase) is discussed.

  17. Biochemical characterization of mutants in the active site residues of the β-galactosidase enzyme of Bacillus circulans ATCC 31382

    OpenAIRE

    Bultema, Jelle B; Bas J.H. Kuipers; Lubbert Dijkhuizen

    2014-01-01

    The Bacillus circulans ATCC 31382 β-galactosidase (BgaD) is a retaining-type glycosidase of glycoside hydrolase family 2 (GH2). Its commercial enzyme preparation, Biolacta N5, is used for commercial-scale production of galacto-oligosaccharides (GOS). The BgaD active site and catalytic amino acid residues have not been studied. Using bioinformatic routines we identified two putative catalytic glutamates and two highly conserved active site histidines. The site-directed mutants E447N, E532Q, an...

  18. Cytochrome cb-type nitric oxide reductase with cytochrome c oxidase activity from Paracoccus denitrificans ATCC 35512.

    OpenAIRE

    T Fujiwara; Fukumori, Y

    1996-01-01

    A highly active nitric oxide reductase was purified from Paracoccus denitrificans ATCC 35512, formerly named Thiosphaera pantotropha, which was anaerobically cultivated in the presence of nitrate. The enzyme was composed of two subunits with molecular masses of 34 and 15 kDa and contained two hemes b and one heme c per molecule. Copper was not found in the enzyme. The spectral properties suggested that one of the two hemes b and heme c were in six-coordinated low-spin states and another heme ...

  19. Effect of fermentation conditions on the production of hyaluronic acid by Streptococcus zooepidemicus ATCC 39920

    OpenAIRE

    Nicole Caldas Pan; Josiane Alessandra Vignoli; Cristiani Baldo; Hanny Cristina Braga Pereira; Rui Sérgio dos Santos Ferreira Silva; Maria Antonia Pedrine Colabone Celligoi

    2015-01-01

    The production of hyaluronic acid by Streptococcus zooepidemicus ATCC 39920 with varying rates of pH (6.0, 7.0, 8.0), temperature (34; 37; 40°C), agitation (100, 150, 200 rpm), glucose (10, 20, 30 g L-1) and yeast extract concentration (10, 20, 30 g L-1) was evaluated by statistical approaches. The best conditions for the production of hyaluronic acid was pH 8.0, 37°C and 100 rpm in a medium containing 30 g L-1 glucose and yeast extract, for a production of 0.787 g L-1. Temperature, pH and ye...

  20. Cloning and Characterization of a Gene (mspA) Encoding the Major Sheath Protein of Treponema maltophilum ATCC 51939T

    OpenAIRE

    Heuner, Klaus; Choi, Bong-Kyu; Schade, Rüdiger; Moter, Annette; Otto, Albrecht; Göbel, Ulf B.

    1999-01-01

    The major sheath protein-encoding gene (mspA) of the oral spirochete Treponema maltophilum ATCC 51939T was cloned by screening a genomic library with an anti-outer membrane fraction antibody. The mspA gene encodes a precursor protein of 575 amino acids with a predicted molecular mass of 62.3 kDa, including a signal peptide of 19 amino acids. The native MspA formed a heat-modifiable, detergent- and trypsin-stable complex which is associated with the outer membrane. Hybridization with an mspA-s...

  1. Extracellular lipase of Pseudomonas sp. strain ATCC 21808: purification, characterization, crystallization, and preliminary X-ray diffraction data.

    OpenAIRE

    Kordel, M; Hofmann, B; Schomburg, D; Schmid, R. D.

    1991-01-01

    A procedure for the purification of a very hydrophobic lipase from Pseudomonas sp. strain ATCC 21808 was elaborated by avoiding the use of long-chain detergents in view of subsequent crystallization of the enzyme. The purification procedure included chromatography on Q-Sepharose in the presence of n-octyl-beta-D-glucopyranoside, Ca2+ precipitation of fatty acids, and Octyl-Sepharose chromatography. The enzyme was purified 260-fold to a yield of 35% and a specific activity of 3,300 U/mg. The m...

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

    OpenAIRE

    Vengadaramana, A.; Vasanthy, A.; Balakumar, S

    2012-01-01

    Aims: The effect of carbohydrate and amino acids on the production of a-amylase by Bacillus licheniformis ATCC 6346 was investigated. Methodology and results: To find out the influence of carbohydrate the total carbohydrate content of the medium containing different concentration (2-18 g/L) of defatted seed cake powder of sesamum and mustard containing medium was kept constant by the addition of soluble starch separately. The highest a-amylase activity obtained in the medium containing 18g/L ...

  3. Comparative genomics of citric-acid producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88

    Energy Technology Data Exchange (ETDEWEB)

    Grigoriev, Igor V.; Baker, Scott E.; Andersen, Mikael R.; Salazar, Margarita P.; Schaap, Peter J.; Vondervoot, Peter J.I. van de; Culley, David; Thykaer, Jette; Frisvad, Jens C.; Nielsen, Kristen F.; Albang, Richard; Albermann, Kaj; Berka, Randy M.; Braus, Gerhard H.; Braus-Stromeyer, Susanna A.; Corrochano, Luis M.; Dai, Ziyu; Dijck, Piet W.M. van; Hofmann, Gerald; Lasure, Linda L.; Magnusson, Jon K.; Meijer, Susan L.; Nielsen, Jakob B.; Nielsen, Michael L.; Ooyen, Albert J.J. van; Panther, Kathyrn S.; Pel, Herman J.; Poulsen, Lars; Samson, Rob A.; Stam, Hen; Tsang, Adrian; Brink, Johannes M. van den; Atkins, Alex; Aerts, Andrea; Shapiro, Harris; Pangilinan, Jasmyn; Salamov, Asaf; Lou, Yigong; Lindquist, Erika; Lucas, Susan; Grimwood, Jane; Kubicek, Christian P.; Martinez, Diego; Peij, Noel N.M.E. van; Roubos, Johannes A.; Nielsen, Jens

    2011-04-28

    The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzyme-producing A. niger strain (CBS 513.88) has already been sequenced, the versatility and diversity of this species compels additional exploration. We therefore undertook whole genome sequencing of the acidogenic A. niger wild type strain (ATCC 1015), and produced a genome sequence of very high quality. Only 15 gaps are present in the sequence and half the telomeric regions have been elucidated. Moreover, sequence information from ATCC 1015 was utilized to improve the genome sequence of CBS 513.88. Chromosome-level comparisons uncovered several genome rearrangements, deletions, a clear case of strain-specific horizontal gene transfer, and identification of 0.8 megabase of novel sequence. Single nucleotide polymorphisms per kilobase (SNPs/kb) between the two strains were found to be exceptionally high (average: 7.8, maximum: 160 SNPs/kb). High variation within the species was confirmed with exo-metabolite profiling and phylogenetics. Detailed lists of alleles were generated, and genotypic differences were observed to accumulate in metabolic pathways essential to acid production and protein synthesis. A transcriptome analysis revealed up-regulation of the electron transport chain, specifically the alternative oxidative pathway in ATCC 1015, while CBS 513.88 showed significant up-regulation of genes relevant to glucoamylase A production, such as tRNA-synthases and protein transporters. Our results and datasets from this integrative systems biology analysis resulted in a snapshot of fungal evolution and will support further optimization of cell factories based on filamentous fungi.[Supplemental materials (10 figures, three text documents and 16 tables) have been made available

  4. Inhibition of Listeria monocytogenes ATCC 19115 on ham steak by tea bioactive compounds incorporated into chitosan-coated plastic films

    Directory of Open Access Journals (Sweden)

    Vodnar Dan C

    2012-07-01

    Full Text Available Abstract Background The consumer demands for better quality and safety of food products have given rise to the development and implementation of edible films. The use of antimicrobial films can be a promising tool for controlling L. monocytogenes on ready to eat products. The aim of this study was to develop effective antimicrobial films incorporating bioactive compounds from green and black teas into chitosan, for controlling L. monocytogenes ATCC 19115 on vacuum-packaged ham steak. The effectiveness of these antimicrobial films was evaluated at room temperature (20°C for 10 days and at refrigerated temperature (4°C for 8 weeks. Results The HPLC results clearly show that relative concentrations of catechins and caffeine in green tea ranked EGCG>EGC>CAF>ECG>EC>C while in black tea extracts ranked CAF>EGCG>ECG>EGC>EC>C. The chitosan-coated plastic films incorporating green tea and black tea extracts shows specific markers identified by FTIR. Incorporating natural extracts into chitosan showed that the growth of L monocytogenes ATCC 19115 was inhibited. The efficacy of antimicrobial effect of tea extracts incorporated into chitosan-coated plastic film was dose dependent. However, chitosan-coated films without addition of tea extracts did not inhibit the growth of L. monocytogenes ATCC 19115. Chitosan-coated plastic films incorporating 4% Green tea extract was the most effective antimicrobial, reducing the initial counts from 3.2 to 2.65 log CFU/cm2 during room temperature storage and from 3.2 to 1–1.5 log CFU/cm2 during refrigerated storage. Conclusions Incorporation of tea extracts into the chitosan-coated films considerably enhanced their effectiveness against L. monocytogenes ATCC 19115. 4% Green tea incorporated into chitosan-coated plastic film had a better antilisterial effect than 2% green tea or 2% and 4% black tea. Data from this study would provide new formulation options for developing antimicrobial packaging films using tea

  5. Evaluation of probiotic properties of Pediococcus acidilactici B14 in association with Lactobacillus acidophilus ATCC 4356 for application in a soy based aerated symbiotic dessert

    Directory of Open Access Journals (Sweden)

    Maria Carolina de Oliveira Ribeiro

    2014-10-01

    Full Text Available The aim of this study was to evaluate the probiotic properties of Pediococcus acidilactici B14 and to study its resistance in the gastrointestinal system when combined with Lactobacillus acidophilus ATCC 4356 and used in a potentially symbiotic aerated soy based dessert. P. acidilactici B14 showed some important probiotic characteristics such as survival rate of 45.9% at pH 2.5; 72.4% in 0.3% bile salts and 95.8% after gastrointestinal transit at pH 4.0. Tolerance against the antibiotics cephalexin, neomycin, vancomycin, cefotaxime and penicillin G was also observed. The strain inhibited antagonism against the following cultures: Escherichia coli ATCC 25922, Bacillus cereus ATCC 33018, Staphylococcus aureus ATCC 6538P and Salmonella sp. The mixed culture of P. acidilactici B14 with L. acidophilus ATCC 4356 showed a survival rate of 92.4% after the passage through the gastrointestinal system at pH 4.0. Furthermore, in the presence of the food matrix, an average increase in cell viability, after being subjected to the gastrointestinal system of 9.9% at pH 2.0 and 6.1% at pH 4.0, was observed. This characterized the adequacy of the associated culture as probiotic in the development of a functional food such as soy based aerated symbiotic dessert.

  6. Strain improvement of Sporolactobacillus inulinus ATCC 15538 for acid tolerance and production of D-lactic acid by genome shuffling.

    Science.gov (United States)

    Zheng, Huijie; Gong, Jixian; Chen, Tao; Chen, Xun; Zhao, Xueming

    2010-02-01

    Improvement of acid tolerance and production of D-lactic acid by Sporolactobacillus inulinus ATCC 15538 was performed by using recursive protoplast fusion in a genome shuffling format. The starting population was generated by ultraviolet irradiation, diethyl sulfate mutagenesis, and pH-gradient filter and then, subjected for the recursive protoplast fusion. The concentration of lysozyme, time, and temperature for enzyme treatment were optimized by response surface methodology based on the central composite design. Based on contour plots and variance analysis, the model predicted a maximum Y (multiply protoplasts formation ratio by protoplasts regeneration ratio), 60.4%, and the corresponding above used values were 7.75 mg/ml lysozyme, 1.59 h, and 38 degrees C. A pH-5-resistant recombinant, F3-4, was obtained after three rounds of genome shuffling and its production of D-lactic acid reached 93.4 g/l in a 5 L bioreactor, which was increased by 39.8% and 119% in comparison with that of UV generated strain and the original strain S. inulinus ATCC 15538, respectively. The subculture experiments indicated that F3-4 was genetically stable. PMID:19777227

  7. Inactivation of Listeria monocytogenes ATCC 7644 on fresh-cut tomato using nisin in combinations with organic salts.

    Science.gov (United States)

    Oladunjoye, Adebola O; Singh, Suren; Ijabadeniyi, Oluwatosin A

    2016-01-01

    The inhibition of Listeria monocytogenes ATCC 7644 on fresh-cut tomato was investigated using nisin alone, and in combinations with organic salts. Nisin at a concentration of 5000UI/mL was introduced alone or in combination with an organic salt (sodium citrate or sodium acetate each at 3 and 5g/100mL each) on fresh-cut tomato previously inoculated with 10(8)CFU/mL of L. monocytogenes ATCC 7644. Chlorine at 200ppm was used as a control. The inoculated samples were incubated at different temperatures (4, 10 and 25°C) and examined at 0, 24, 48 and 72h. The effects of the antimicrobial treatments on quality parameters of tomato (pH, soluble solids, titratable acidity and vitamin C) were also evaluated, and colour parameters were observed at the lowest storage temperature for 10 days. Both nisin and the organic salts inhibited growth of L. monocytogenes, but the combinations of two compounds were more effective. The nisin-sodium citrate (5%) combination was significantly (p≤0.05) effective, while chlorine was least effective against L. monocytogenes. The quality parameters were substantially retained, especially at 4°C, suggesting good shelf stability at a low temperature. These results substantiate the use of the cheap and eco-friendly approach to reducing this pathogen of health concern in common fresh produce. PMID:27261167

  8. Cytochrome cb-type nitric oxide reductase with cytochrome c oxidase activity from Paracoccus denitrificans ATCC 35512.

    Science.gov (United States)

    Fujiwara, T; Fukumori, Y

    1996-04-01

    A highly active nitric oxide reductase was purified from Paracoccus denitrificans ATCC 35512, formerly named Thiosphaera pantotropha, which was anaerobically cultivated in the presence of nitrate. The enzyme was composed of two subunits with molecular masses of 34 and 15 kDa and contained two hemes b and one heme c per molecule. Copper was not found in the enzyme. The spectral properties suggested that one of the two hemes b and heme c were in six-coordinated low-spin states and another heme b was in a five-coordinated high-spin state and reacted with carbon monoxide. The enzyme showed high cytochrome c-nitric oxide oxidoreductase activity and formed nitrous oxide from nitric oxide with the expected stoichiometry when P. denitrificans ATCC 35512 ferrocytochrome c-550 was used as the electron donor. The V max and Km values for nitric oxide were 84 micromol of nitric oxide per min/mg of protein and 0.25 microM, respectively. Furthermore, the enzyme showed ferrocytochrome c-550-O2 oxidoreductase activity with a V max of 8.4 micromol of O2 per min/mg of protein and a Km value of 0.9 mM. Both activities were 50% inhibited by about 0.3 mM KCN. PMID:8606159

  9. Complete structure of the cell surface polysaccharide of Streptococcus oralis ATCC 10557: A receptor for lectin-mediated interbacterial adherence

    Energy Technology Data Exchange (ETDEWEB)

    Abeygunawardana, C.; Bush, C.A. (Univ. of Maryland, Baltimore (United States)); Cisar, J.O. (National Inst. of Dental Research, Bethesda, MD (United States))

    1991-07-02

    Lectin-carbohydrate binding is known to play an important role in a number of different cell-cell interactions including those between certain species of oral streptococci and actinomyces that colonize teeth. The cell wall polysaccharides of Streptococcus oralis ATCC 10557, S. oralis 34, and Streptococcus mitis J22, although not identical antigenically, each function as a receptor molecule for the galactose and N-acetylgalactosamine reactive fimbrial lectins of Actinomyces viscosus and Actinomyces naeslundii. Carbohydrate analysis of the receptor polysaccharide isolated from S. oralis ATCC 10557 shows galactose (3 mol), glucose (1 mol), GalNAc (1 mol), and rhamnose (1 mol). {sup 1}H NMR spectra of the polysaccharide show that is partially O-acetylated. Analysis of the {sup 1}H NMR spectrum of the de-O-acetylated polysaccharide shows that it is composed of repeating subunits containing six monosaccharides and that the subunits are joined by a phosphodiester linkage. The {sup 1}H and {sup 13}C NMR spectra were completely assigned by two-dimensional homonuclear correlation methods and by {sup 1}H-detected heteronuclear multiple-quantum correlation ({sup 1}H({sup 13}C)HMQC). The complete {sup 1}H and {sup 13}C assignment of the native polysaccharide was carried out by the same techniques augmented by a {sup 13}C-coupled hybrid HMQC-COSY method, which is shown to be especially useful for carbohydrates in which strong coupling and overlapping peaks in the {sup 1}H spectrum pose difficulties.

  10. Free and immobilized Lactobacillus casei ATCC 393 on whey protein as starter cultures for probiotic Feta-type cheese production.

    Science.gov (United States)

    Dimitrellou, Dimitra; Kandylis, Panagiotis; Sidira, Marianthi; Koutinas, Athanasios A; Kourkoutas, Yiannis

    2014-01-01

    The use of free and immobilized Lactobacillus casei ATCC 393 on whey protein as starter culture in probiotic Feta-type cheese production was evaluated. The probiotic cultures resulted in significantly higher acidity; lower pH; reduced counts of coliforms, enterobacteria, and staphylococci; and improved quality characteristics compared with cheese with no culture. Microbiological and strain-specific multiplex PCR analysis showed that both free and immobilized L. casei ATCC 393 were detected in the novel products at levels required for conferring a probiotic effect at the end of the ripening. The effect of starter culture on production of volatile compounds was investigated by the solid-phase microextraction gas chromatography-mass spectrometry analysis technique. The immobilized cells resulted in an improved profile of aroma-related compounds and the overall high quality of the novel products was ascertained by the preliminary sensory test. Finally, the high added value produced by exploitation of whey, which is an extremely polluting industrial waste, was highlighted and assessed. PMID:24931523

  11. Antiserum against Raoultella terrigena ATCC 33257 identifies a large number of Raoultella and Klebsiella clinical isolates as serotype O12.

    Science.gov (United States)

    Mertens, Katja; Müller-Loennies, Sven; Stengel, Petra; Podschun, Rainer; Hansen, Dennis S; Mamat, Uwe

    2010-12-01

    Raoultella terrigena ATCC 33257, recently reclassified from the genus Klebsiella, is a drinking water isolate and belongs to a large group of non-typeable Klebsiella and Raoultella strains. Using an O-antiserum against a capsule-deficient mutant of this strain, we could show a high prevalence (10.5%) of the R. terrigena O-serotype among non-typeable, clinical Klebsiella and Raoultella isolates. We observed a strong serological cross-reaction with the K. pneumoniae O12 reference strain, indicating that a large percentage of these non-typeable strains may belong to the O12 serotype, although these are currently not detectable by the K. pneumoniae O12 reference antiserum in use. Therefore, we analyzed the O-polysaccharide (O-PS) structure and genetic organization of the wb gene cluster of R. terrigena ATCC 33257, and both confirmed a close relation of R. terrigena and K. pneumoniae O12. The two strains possess an identical O-PS, lipopolysaccharide core structure, and genetic organization of the wb gene cluster. Heterologous expression of the R. terrigena wb gene cluster in Escherichia coli K-12 resulted in the WecA-dependent synthesis of an O-PS reactive with the K. pneumoniae O12 antiserum. The serological data presented here suggest a higher prevalence of the O12-serotype among Klebsiella and Raoultella isolates than generally assumed.

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

    Directory of Open Access Journals (Sweden)

    Vengadaramana, A.

    2012-01-01

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

  13. Effect of nutrient media on photobiological hydrogen production by Anabaena variabilis ATCC 29413

    Energy Technology Data Exchange (ETDEWEB)

    Berberoglu, Halil; Pilon, Laurent [Mechanical and Aerospace Engineering Department, Henry Samueli School of Engineering and Applied Science, University of California Los Angeles, Los Angeles, CA 90095 (United States); Jay, Jenny [Civil and Environmental Engineering Department, Henry Samueli School of Engineering and Applied Science, University of California Los Angeles, Los Angeles, CA 90095 (United States)

    2008-02-15

    This study reports a factor 5.5 increase in hydrogen production by Anabaena variabilis ATCC 29413 using Allen-Arnon medium compared with BG-11 and BG-11{sub 0} media. The results were obtained with a flat panel photobioreactor made of acrylic and operated in two stages at 30 C. Stage 1 aims at converting carbon dioxide into biomass by photosynthesis while Stage 2 aims at producing hydrogen. During Stage 1, the photobioreactor was irradiated with 65{mu}mol/m{sup 2}/s (14W/m{sup 2}) of light and sparged with a mixture of air (95% by volume) and carbon dioxide (5% by volume). During Stage 2, irradiance was increased to 150{mu}mol/m{sup 2}/s (32W/m{sup 2}) and the photobioreactor was sparged with pure argon. The parameters continuously monitored were (1) the cyanobacteria concentration, (2) the pH, (3) the dissolved oxygen concentration, (4) the nitrate and (5) the ammonia concentrations in the medium, and (6) the hydrogen concentration in the effluent gas. The three media BG-11, BG-11{sub 0}, and Allen-Arnon were tested under otherwise similar conditions. The maximum cyanobacteria concentrations during Stage 2 were 1.10 and 1.17kg drycell/m{sup 3} with BG-11 and Allen-Arnon media, respectively, while it could not exceed 0.76kg drycell/m{sup 3} with medium BG-11{sub 0}. Moreover, the heterocyst frequency was 5%, 4%, and 9% for A.variabilis grown in BG-11, BG-11{sub 0}, and Allen-Arnon media. The average specific hydrogen production rates were about 8.0 x 10{sup -5} and 7.2 x 10{sup -5}kgH{sub 2}/kgdrycell/h (1 and 0.9LH{sub 2}/kgdrycell/h at 1 atm and 30 {sup o}C) in media BG-11 and BG-11{sub 0}, respectively. In contrast, it was about 4.5 x 10{sup -4}kgH{sub 2}/kgdrycell/h (5.6LH{sub 2}/kgdrycell/h at 1 atm and 30 {sup o}C) in Allen-Arnon medium. The maximum light to hydrogen energy conversion efficiencies achieved were 0.26%, 0.16%, and 1.32% for BG-11, BG-11{sub 0}, and Allen-Arnon media, respectively. The larger heterocyst frequency, specific hydrogen production

  14. Microbioligcal Transformation of Bavachinin by Penicillium Raistrickii ATCC10490%雷斯青霉ATCC10490对补骨脂二氢黄酮甲醚转化的研究

    Institute of Scientific and Technical Information of China (English)

    梁奇坤; 王家明; 申雁冰; 陈曦; 孙华; 王敏

    2012-01-01

    The biotransformation of bavachinin with Penicillium raistrickii ATCC 10490 gave one compound with the name of BC-PC-4. This product was purified from broth culture supernatants by silica gel column chromatography. Bavachinin and BC-PC-4 were evaluated by the cell viability test through the way of MTT assay with the control of taxol. The compound BC-PC-4 exhibited the higher tumor cytotoxicity than that of bavachinin towards MCF-7 cell, with the IC50 value of 9. 31 μmol/L, which is 7. 04 μmol/L lower than that of bavachinin. On the other hand,the compound BC-PC-4 showed the ability of proliferating of HUVEC cells.%研究了雷斯青霉(Penicillium raistrickii ATCC 10490对补骨脂二氢黄酮甲醚生物的微生物转化,通过硅胶柱分离等技术得到转化产物BC-PC-4的纯品.在细胞水平上分析了该转化产物的抗肿瘤活性,结果发现产物BC-PC-4对细胞MCF-7的抗肿瘤活性IC50值为9.31 μmol/L,较底物补骨脂二氢黄酮甲醚降低了7.04 μmol/L,且对正常细胞HUVEC有促进增殖的作用.

  15. Growth and enzyme production of Cellulomonas sp. ATCC 21399 on microcrystalline cellulose. Effects of increasing concentration of a mineral medium

    Energy Technology Data Exchange (ETDEWEB)

    Poulsen, O.M.; Petersen, L.W.

    1989-05-01

    The kinetics and production of different extracellular enzyme activities were studied during growth of Cellulomonas sp. ATCC 21399 on 2% Avicel with different concentrations of M9 mineral medium. The lag phase and the doubling time increased with increasing ionic strength of the medium. The highest cell density was obtained during growth at 5xM9 mineral medium and Cellulomonas grew well at this high salinity. The enzyme activities against carboxymethylcellulose and xylan increased with increasing concentration of M9 medium up to 5xM9. By contrast, activities against microcrystalline cellulose (Avicel), galactomannan and amylose decreased with increasing concentration of M9 medium. The extracellular proteinase activity increased with increasing concentration of M9 medium, and it is possible that the lability of the cellulolytic and amylolytic enzymes may be due to their susceptibility to proteolytic inactivation by the extracellular proteinases.

  16. AcsA-AcsB: The core of the cellulose synthase complex from Gluconacetobacter hansenii ATCC23769.

    Science.gov (United States)

    McManus, John B; Deng, Ying; Nagachar, Nivedita; Kao, Teh-hui; Tien, Ming

    2016-01-01

    The gram-negative bacterium, Gluconacetobacter hansenii, produces cellulose of exceptionally high crystallinity in comparison to the cellulose of higher plants. This bacterial cellulose is synthesized and extruded into the extracellular medium by the cellulose synthase complex (CSC). The catalytic component of this complex is encoded by the gene AcsAB. However, several other genes are known to encode proteins critical to cellulose synthesis and are likely components of the bacterial CSC. We have purified an active heterodimer AcsA-AcsB from G. hansenii ATCC23769 to homogeneity by two different methods. With the purified protein, we have determined how it is post-translationally processed, forming the active heterodimer AcsA-AcsB. Additionally, we have performed steady-state kinetic studies on the AcsA-AcsB complex. Finally through mutagenesis studies, we have explored the roles of the postulated CSC proteins AcsC, AcsD, and CcpAx.

  17. Supporting data for comparative proteomic analysis of Listeria monocytogenes ATCC 7644 exposed to a sublethal concentration of nisin

    Directory of Open Access Journals (Sweden)

    Kendi Nishino Miyamoto

    2015-06-01

    Full Text Available Here we provide the LC–MS/MS data from a comparative analysis of Listeria monocytogenes ATCC 7644 treated and non-treated with a sublethal concentration of nisin (10−3 mg/mL. Protein samples were analyzed by multidimensional protein identification technology (MudPIT approach, in an off-line configuration. The raw MS/MS data allowed the detection of 49,591 spectra which resulted in 576 protein identifications. After Scaffold validation, 179 proteins were identified with high confidence. A label-free quantitative analysis based of normalized spectral abundance factor (NSAF was used and 13 proteins were found differentially expressed between nisin-treated and non-treated cells. Gene ontology analysis of differentially expressed proteins revealed that most of them are correlated to metabolic process, oxidative stress response mechanisms and molecular binding. A detailed analysis and discussion of these data may be found in Miyamoto et al. [1].

  18. Development of an Unnatural Amino Acid Incorporation System in the Actinobacterial Natural Product Producer Streptomyces venezuelae ATCC 15439.

    Science.gov (United States)

    He, Jingxuan; Van Treeck, Briana; Nguyen, Han B; Melançon, Charles E

    2016-02-19

    Many Actinobacteria, most notably Streptomyces, produce structurally diverse bioactive natural products, including ribosomally synthesized peptides, by multistep enzymatic pathways. The use of site-specific genetic incorporation of unnatural amino acids to investigate and manipulate the functions of natural product biosynthetic enzymes, enzyme complexes, and ribosomally derived peptides in these organisms would have important implications for drug discovery and development efforts. Here, we have designed, constructed, and optimized unnatural amino acid systems capable of incorporating p-iodo-l-phenylalanine and p-azido-l-phenylalanine site-specifically into proteins in the model natural product producer Streptomyces venezuelae ATCC 15439. We observed notable differences in the fidelity and efficiency of these systems between S. venezuelae and previously used hosts. Our findings serve as a foundation for using an expanded genetic code in Streptomyces to address questions related to natural product biosynthesis and mechanism of action that are relevant to drug discovery and development. PMID:26562751

  19. Characterization and replication mode determination of the minimal replicon of Tetragenococcus halophila ATCC33315 plasmid pUCL287.

    Science.gov (United States)

    Benachour, A; Frère, J; Boutibonnes, P; Auffray, Y

    1995-01-01

    pUCL287 is a cryptic plasmid of Tetragenococcus halophila (formerly Pediococcus halophilus) ATCC33315 of relatively small size (8.7 kb). Its minimal replicon was located on a 1235 bp MamI-EcoRI fragment. This minimal replicon contains a non-translated region, followed by a gene encoding a putative 311 amino acid protein. Deletion experiments showed that the non-translated region corresponds to the replication origin. Determination of the replication mode was carried out in Enterococcus faecalis JH2-2 harboring pUCL287 minimal replicon. The replicating intermediates detected revealed that pUCL287 minimal replicon follows a bidirectional theta replicating mode. PMID:8824766

  20. Supporting data for comparative proteomic analysis of Listeria monocytogenes ATCC 7644 exposed to a sublethal concentration of nisin

    Science.gov (United States)

    Miyamoto, Kendi Nishino; Mariante Monteiro, Karina; da Silva Caumo, Karin; Rodrigues Lorenzatto, Karina; Bunselmeyer Ferreira, Henrique; Brandelli, Adriano

    2015-01-01

    Here we provide the LC–MS/MS data from a comparative analysis of Listeria monocytogenes ATCC 7644 treated and non-treated with a sublethal concentration of nisin (10−3 mg/mL). Protein samples were analyzed by multidimensional protein identification technology (MudPIT) approach, in an off-line configuration. The raw MS/MS data allowed the detection of 49,591 spectra which resulted in 576 protein identifications. After Scaffold validation, 179 proteins were identified with high confidence. A label-free quantitative analysis based of normalized spectral abundance factor (NSAF) was used and 13 proteins were found differentially expressed between nisin-treated and non-treated cells. Gene ontology analysis of differentially expressed proteins revealed that most of them are correlated to metabolic process, oxidative stress response mechanisms and molecular binding. A detailed analysis and discussion of these data may be found in Miyamoto et al. [1]. PMID:26217729

  1. Utilisation of vegetable oils in the production of lovastatin by Aspergillus terreus ATCC 20542 in submerged cultivation

    Directory of Open Access Journals (Sweden)

    Pattana Sripalakit

    2011-07-01

    Full Text Available The effect of vegetable oils as a supplementary carbon source during the production of lovastatin by Aspergillus terreus ATCC 20542 in submerged culture was investigated. The six vegetable oils tested were sesame oil, sunflower oil, soya bean oil, corn oil, palm oil and olive oil. Lovastatin concentration and biomass were measured. Lovastatin production was higher in several oil-containing media compared to control medium. In particular, palm oil and soya bean oil significantly improved lovastatin production. Yields with palm oil and soya bean oil were 4.5- and 1.4-fold higher respectively, compared with control. Sesame oil and corn oil, however, had a negative effect on lovastatin production. Biomass was proportional to vegetable oil concentration, but an excessive vegetable oil concentration resulted in a lower yield of lovastatin. Thus, some vegetable oils appear to be excellent adjuvants for improving efficiency of lovastatin production.

  2. AcsA-AcsB: The core of the cellulose synthase complex from Gluconacetobacter hansenii ATCC23769.

    Science.gov (United States)

    McManus, John B; Deng, Ying; Nagachar, Nivedita; Kao, Teh-hui; Tien, Ming

    2016-01-01

    The gram-negative bacterium, Gluconacetobacter hansenii, produces cellulose of exceptionally high crystallinity in comparison to the cellulose of higher plants. This bacterial cellulose is synthesized and extruded into the extracellular medium by the cellulose synthase complex (CSC). The catalytic component of this complex is encoded by the gene AcsAB. However, several other genes are known to encode proteins critical to cellulose synthesis and are likely components of the bacterial CSC. We have purified an active heterodimer AcsA-AcsB from G. hansenii ATCC23769 to homogeneity by two different methods. With the purified protein, we have determined how it is post-translationally processed, forming the active heterodimer AcsA-AcsB. Additionally, we have performed steady-state kinetic studies on the AcsA-AcsB complex. Finally through mutagenesis studies, we have explored the roles of the postulated CSC proteins AcsC, AcsD, and CcpAx. PMID:26672449

  3. Erythromycin and azithromycin transport into Haemophilus influenzae ATCC 19418 under conditions of depressed proton motive force (delta mu H)

    Energy Technology Data Exchange (ETDEWEB)

    Capobianco, J.O.; Goldman, R.C. (Abbott Laboratories, IL (USA))

    1990-09-01

    The effect of collapsing the electrochemical proton gradient (delta mu H) on ({sup 3}H)erythromycin and ({sup 14}C)azithromycin transport in Haemophilus influenzae ATCC 19418 was studied. The proton gradient and membrane potential were determined from the distribution of (2-{sup 14}C)dimethadione and rubidium-86, respectively. delta mu H was reduced from 124 to 3 mV in EDTA-valinomycin-treated cells at 22{degrees}C with 150 mM KCl and 0.1 mM carbonyl cyanide m-chlorophenylhydrazone. During the collapse of delta mu H, macrolide uptake increased. Erythromycin efflux studies strongly suggested that this increase was not due to an energy-dependent efflux pump but was likely due to increased outer membrane permeability. These data indicated that macrolide entry was not a delta mu H-driven active transport process but rather a passive diffusion process.

  4. Influence of sporulation medium composition on transcription of ger operons and the germination response of spores of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Hornstra, L.M.; Vries, de Y.P.; Vos, de W.M.; Abee, T.

    2006-01-01

    Bacillus cereus ATCC 14579 endospores were produced in Y1 medium, a nutrient-rich, chemically defined sporulation medium, and in modified G medium, containing low amounts of nutrients. The average transcription level of the seven ger operons per cell was 3.5 times higher in Y1 medium, and the spores

  5. Genome sequence of Victivallis vadensis ATCC BAA-548, an anaerobic bacterium from the phylum Lentisphaerae, isolated from the human gastro-intestinal tract

    Energy Technology Data Exchange (ETDEWEB)

    Van Passel, Mark W.J. [Wageningen University and Research Centre, The Netherlands; Kant, Ravi [University of Helsinki; Palva, Airi [University of Helsinki; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Bruce, David [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Davenport, Karen W. [Los Alamos National Laboratory (LANL); Sims, David [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Han, Cliff [Los Alamos National Laboratory (LANL); Larimer, Frank W [ORNL; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Ovchinnikova, Galina [U.S. Department of Energy, Joint Genome Institute; Richardson, Paul [U.S. Department of Energy, Joint Genome Institute; De Vos, Willem M. [Wageningen University and Research Centre, The Netherlands; Smidt, Hauke [Wageningen University and Research Centre, The Netherlands; Zoetendal, Erwin G. [Wageningen University and Research Centre, The Netherlands

    2011-01-01

    Victivallis vadensis ATCC BAA-548 represents the first cultured representative from the novel phylum Lentisphaerae, a deep-branching bacterial lineage. Few cultured bacteria from this phylum are known, and V. vadensis therefore represents an important organism for evolutionary studies. V. vadensis is a strictly anaerobic sugar-fermenting isolate from the human gastro-intestinal tract.

  6. 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

  7. 40 CFR 180.1102 - Trichoderma harzianum KRL-AG2 (ATCC #20847) strain T-22; exemption from requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Trichoderma harzianum KRL-AG2 (ATCC... PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1102 Trichoderma harzianum KRL-AG2... of a tolerance is established for residues of the biofungicide Trichoderma harzianum KRL-AG2...

  8. Complete Genome Sequence of a Gluconacetobacter hansenii ATCC 23769 Isolate, AY201, Producer of Bacterial Cellulose and Important Model Organism for the Study of Cellulose Biosynthesis.

    Science.gov (United States)

    Pfeffer, Sarah; Mehta, Kalpa; Brown, R Malcolm

    2016-08-11

    The cellulose producer and model organism used for the study of cellulose biosynthesis, Gluconacetobacter hansenii AY201, is a variant of G. hansenii ATCC 23769. We report here the complete nucleotide sequence of G. hansenii AY201, information which may be utilized to further the research into understanding the genes necessary for cellulose biosynthesis.

  9. Reduced hybrid cluster proteins (HCP) from Desulfovibrio desulfuricans ATCC 27774 and Desulfovibrio vulgaris (Hildenborough): X-ray structures at high resolution using synchrotron radiation

    NARCIS (Netherlands)

    Aragao, D.; Macedo, S.; Mitchell, E.P.; Romao, C.V.; Liu, M.Y.; Frazao, C.; Saraiva, L.M.; Xavier, A.V.; LeGall, J.; Dongen, van W.M.A.M.; Hagen, W.R.; Teixeira, M.; Carrondo, M.A.; Lindley, P.

    2003-01-01

    The hybrid cluster proteins from the sulfate reducing bacteria Desulfovibrio desulfuricans ATCC 27774 (Dd) and Desulfovibrio vulgaris strain Hildenborough(Dv) have been isolated and crystallized anaerobically. In each case, the protein has been reduced with dithionite and the crystal structure of th

  10. Expression of cbsA encoding the collagen-binding S-protein of Lactobacillus crispatus JCM5810 in Lactobacillus casei ATCC 393T

    NARCIS (Netherlands)

    Martínez, B.; Sillanpää, J.; Smit, E.; Korhonen, T.K.; Pouwels, P.H.

    2000-01-01

    The cbsA gene encoding the collagen-binding S-layer protein of Lactobacillus crispatus JCM5810 was expressed in L. casei ATCC 393T. The S-protein was not retained on the surface of the recombinant bacteria but was secreted into the medium. By translational fusion of CbsA to the cell wall sorting sig

  11. No evidence of harms of probiotic Lactobacillus rhamnosus GG ATCC 53103 in healthy elderly-a Phase I Open Label Study to assess safety, tolerability and cytokine responses

    Science.gov (United States)

    Although Lactobacillus rhamnosus GG ATCC 53103 (LGG) has been consumed since the mid 1990s by between 2 and 5 million people daily, the scientific literature lacks rigorous clinical trials that describe the potential harms of LGG, particularly in the elderly. The primary objective of this open label...

  12. Complete Genome Sequence of a Gluconacetobacter hansenii ATCC 23769 Isolate, AY201, Producer of Bacterial Cellulose and Important Model Organism for the Study of Cellulose Biosynthesis

    Science.gov (United States)

    Mehta, Kalpa

    2016-01-01

    The cellulose producer and model organism used for the study of cellulose biosynthesis, Gluconacetobacter hansenii AY201, is a variant of G. hansenii ATCC 23769. We report here the complete nucleotide sequence of G. hansenii AY201, information which may be utilized to further the research into understanding the genes necessary for cellulose biosynthesis. PMID:27516506

  13. Draft genome sequence of the extremely acidophilic biomining bacterium Acidithiobacillus thiooxidans ATCC 19377 provides insights into the evolution of the Acidithiobacillus genus.

    Science.gov (United States)

    Valdes, Jorge; Ossandon, Francisco; Quatrini, Raquel; Dopson, Mark; Holmes, David S

    2011-12-01

    Acidithiobacillus thiooxidans is a mesophilic, extremely acidophilic, chemolithoautotrophic gammaproteobacterium that derives energy from the oxidation of sulfur and inorganic sulfur compounds. Here we present the draft genome sequence of A. thiooxidans ATCC 19377, which has allowed the identification of genes for survival and colonization of extremely acidic environments.

  14. Draft Genome Sequence of the Extremely Acidophilic Biomining Bacterium Acidithiobacillus thiooxidans ATCC 19377 Provides Insights into the Evolution of the Acidithiobacillus Genus

    OpenAIRE

    Valdes, Jorge; Ossandon, Francisco; Quatrini, Raquel; Dopson, Mark; Holmes, David S

    2011-01-01

    Acidithiobacillus thiooxidans is a mesophilic, extremely acidophilic, chemolithoautotrophic gammaproteobacterium that derives energy from the oxidation of sulfur and inorganic sulfur compounds. Here we present the draft genome sequence of A. thiooxidans ATCC 19377, which has allowed the identification of genes for survival and colonization of extremely acidic environments.

  15. Complete Genome Sequence of a Gluconacetobacter hansenii ATCC 23769 Isolate, AY201, Producer of Bacterial Cellulose and Important Model Organism for the Study of Cellulose Biosynthesis.

    Science.gov (United States)

    Pfeffer, Sarah; Mehta, Kalpa; Brown, R Malcolm

    2016-01-01

    The cellulose producer and model organism used for the study of cellulose biosynthesis, Gluconacetobacter hansenii AY201, is a variant of G. hansenii ATCC 23769. We report here the complete nucleotide sequence of G. hansenii AY201, information which may be utilized to further the research into understanding the genes necessary for cellulose biosynthesis. PMID:27516506

  16. gerR, a novel ger operon involved in L-alanine- and inosine-initiated germination of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

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

    2005-01-01

    Bacillus cereus endospores germinate in response to particular nutrients. Spores are able to sense these nutrients in the environment by receptors encoded by the gerA family of operons. Analysis of the Bacillus cereus ATCC 14579 genome revealed seven gerA family homologues. Using a transposon Tn917-

  17. Characterization of the major dehydrogenase related to d-lactic acid synthesis in Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293.

    Science.gov (United States)

    Li, Ling; Eom, Hyun-Ju; Park, Jung-Mi; Seo, Eunyoung; Ahn, Ji Eun; Kim, Tae-Jip; Kim, Jeong Hwan; Han, Nam Soo

    2012-10-10

    Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293 is a lactic acid bacterium that converts pyruvate mainly to d-(-)-lactic acid by using d-(-)-lactate dehydrogenase (ldhD). The aim of this study was to identify the gene responsible for d-lactic acid formation in this organism and to characterize the enzyme to facilitate the production of optically pure d-lactic acid. A genomic analysis of L. mesenteroides ATCC 8293 revealed that 7 genes encode lactate-related dehydrogenase. According to transcriptomic, proteomic, and phylogenetic analyses, LEUM_1756 was the major gene responsible for the production of d-lactic acid. The LEUM_1756 gene, of 996bp and encoding 332 amino acids (36.5kDa), was cloned and overexpressed in Escherichia coli BL21(DE3) Star from an inducible pET-21a(+) vector. The enzyme was purified by Ni-NTA column chromatography and showed a specific activity of 4450U/mg, significantly higher than those of other previously reported ldhDs. The gel permeation chromatography analysis showed that the purified enzyme exists as tetramers in solution and this was the first report among lactic acid bacteria. The pH and temperature optima were pH 8.0 and 30°C, respectively, for the pyruvate reduction reaction, and pH 11.0 and 20°C, respectively, for the lactate oxidation reaction. The K(m) kinetic parameters for pyruvate and lactate were 0.58mM and 260mM, respectively. In addition, the k(cat) values for pyruvate and lactate were 2900s(-1) and 2280s(-1), respectively. The enzyme was not inhibited by Ca(2+), Co(2+), Cu(2+), Mg(2+), Mn(2+), Na(+), or urea, but was inhibited by 1mM Zn(2+) and 1mM SDS. PMID:22975125

  18. Viability of Escherichia coli ATCC 8739 in Nutrient Broth, Luria-Bertani Broth and Brain Heart Infusion over 11 Weeks

    Directory of Open Access Journals (Sweden)

    Chin How Lee

    2013-01-01

    Full Text Available Abstract:Background: Escherichia coli is a widely studied prokaryotic system. A recent study had demonstrated that reduced growth of E. coli after extended culture in Luria-Bertani broth is a result of depletion of fermentable sugars but able to sustain extended cell culture due to the presence of amino acids, which can be utilized as a carbon source. However, this had not been demonstrated in other media. The study aimed to determine the growth and viability of E. coli ATCC 8739 in 3 different media, Nutrient Broth (NB, Brain Heart Infusion (BHI and Luria-Bertani Broth (LB over 11 weeks.Methods: Growth of E. coli ATCC 8739 was determined by optical density. Viability was determined by serial dilution/spread-plate enumeration. After 11 weeks, the media were exhausted by repeated culture. Glucose was added to the exhausted media to determine whether glucose is the growth-limiting factor.Results: Our results showed that cell density in all 3 media increased to about 1 x 109 cells/ml by the end of week 1, from the inoculation density of 2.67 x 105 cells/ml, peaked at about 1 x 1013 cells/ml at week 4, before declining to about 5 x 107 cells/ml at week 7. Cell density is highly correlated to genomic DNA content (r2 = 0.93 but poorly correlated to optical density (r2< 0.2. Our results also showed that the spent media were able to support further growth after glucose-supplementation.Conclusion: NB, LB and BHI are able to support extended periods of culture and glucose depletion is the likely reason for declining cell growth.

  19. Influence of osmotic stress on the profile and gene expression of surface layer proteins in Lactobacillus acidophilus ATCC 4356.

    Science.gov (United States)

    Palomino, María Mercedes; Waehner, Pablo M; Fina Martin, Joaquina; Ojeda, Paula; Malone, Lucía; Sánchez Rivas, Carmen; Prado Acosta, Mariano; Allievi, Mariana C; Ruzal, Sandra M

    2016-10-01

    In this work, we studied the role of surface layer (S-layer) proteins in the adaptation of Lactobacillus acidophilus ATCC 4356 to the osmotic stress generated by high salt. The amounts of the predominant and the auxiliary S-layer proteins SlpA and SlpX were strongly influenced by the growth phase and high-salt conditions (0.6 M NaCl). Changes in gene expression were also observed as the mRNAs of the slpA and slpX genes increased related to the growth phase and presence of high salt. A growth stage-dependent modification on the S-layer protein profile in response to NaCl was observed: while in control conditions, the auxiliary SlpX protein represented less than 10 % of the total S-layer protein, in high-salt conditions, it increased to almost 40 % in the stationary phase. The increase in S-layer protein synthesis in the stress condition could be a consequence of or a way to counteract the fragility of the cell wall, since a decrease in the cell wall thickness and envelope components (peptidoglycan layer and lipoteichoic acid content) was observed in L. acidophilus when compared to a non-S-layer-producing species such as Lactobacillus casei. Also, the stationary phase and growth in high-salt medium resulted in increased release of S-layer proteins to the supernatant medium. Overall, these findings suggest that pre-growth in high-salt conditions would result in an advantage for the probiotic nature of L. acidophilus ATCC 4356 as the increased amount and release of the S-layer might be appropriate for its antimicrobial capacity. PMID:27376794

  20. Evaluation of molecular techniques for identification and enumeration of Raoultella terrigena ATCC 33257 in water purifier efficacy testing.

    Science.gov (United States)

    Saha, Ratul; Bechanko, Robin; Bestervelt, Lorelle L; Donofrio, Robert S

    2011-09-01

    Raoultella terrigena ATCC 33257, a representative of the coliform group, is commonly used as a challenge organism in water purifier efficacy testing. In addition to being time consuming, traditional culturing techniques and metabolic identification systems (including automated systems) also fail to accurately differentiate this organism from its closely related neighbors belonging to the Enterobacteriaceae group. Molecular-based techniques, such as real-time quantitative polymerase chain reaction (qPCR) and enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting, are preferred methods of detection because of their accuracy, reproducibility, specificity, and sensitivity, along with shorter turnaround time. ERIC-PCR performed with the 1R primer set demonstrated stable unique banding patterns (~800, ~300 bp) for R. terrigena ATCC 33257 different from patterns observed for R. planticola and R. ornithinolytica. The primer pair developed from gyraseA (gyrA) sequence of R. terrigena for the SYBR Green qPCR assay using the AlleleID(®) 7.0 primer probe design software was highly specific and sensitive for the target organism. The sensitivity of the assay was 10(1) colony forming units (CFU)/ml for whole cells and 4.7 fg with genomic DNA. The primer pair was successful in determining the concentration (5.5 ± 0.3 × 10(6) CFU/ml) of R. terrigena from water samples spiked with equal concentration of Escherichia coli and R. terrigena. Based on these results from the ERIC-PCR and the SYBR Green qPCR assay, these molecular techniques can be efficiently used for rapid identification and quantification of R. terrigena during water purifier testing.

  1. Comparison of molecular and biological characteristics of a modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine (ingelvac PRRS MLV), the parent strain of the vaccine (ATCC VR2332), ATCC VR2385, and two recent field isolates of PRRSV.

    Science.gov (United States)

    Opriessnig, T; Halbur, P G; Yoon, K-J; Pogranichniy, R M; Harmon, K M; Evans, R; Key, K F; Pallares, F J; Thomas, P; Meng, X J

    2002-12-01

    The objectives of this study were to compare the molecular and biological characteristics of recent porcine reproductive and respiratory syndrome virus (PRRSV) field isolates to those of a modified live virus (MLV) PRRS vaccine and its parent strain. One hundred seventeen, 4-week-old pigs were randomly assigned to six groups. Group 1 (n = 20) served as sham-inoculated negative controls, group 2 (n = 19) was inoculated with Ingelvac PRRS MLV vaccine, group 3 (n = 20) was inoculated with the parent strain of the vaccine (ATCC VR2332), group 4 (n = 19) was inoculated with vaccine-like PRRSV field isolate 98-38803, group 5 (n = 19) was inoculated with PRRSV field isolate 98-37120, and group 6 (n = 20) was inoculated with known high-virulence PRRSV isolate ATCC VR2385. The levels of severity of gross lung lesions (0 to 100%) among the groups were significantly different at both 10 (P < 0.0001) and 28 days postinoculation (p.i.) (P = 0.002). At 10 days p.i., VR2332 (26.5% +/- 4.64%) and VR2385 (36.4% +/- 6.51%) induced gross lesions of significantly greater severity than 98-38803 (0.0% +/- 0.0%), 98-37120 (0.8% +/- 0.42%), Ingelvac PRRS MLV (0.9% +/- 0.46%), and negative controls (2.3% +/- 1.26%). At 28 days p.i., 98-37120 (17.2% +/- 6.51%) induced gross lesions of significantly greater severity than any of the other viruses. Analyses of the microscopic-interstitial-pneumonia-lesion scores (0 to 6) revealed that VR2332 (2.9 +/- 0.23) and VR2385 (3.1 +/- 0.35) induced significantly more severe lesions at 10 days p.i. At 28 days p.i., VR2385 (2.5 +/- 0.27), VR2332 (2.3 +/- 0.21), 98-38803 (2.6 +/- 0.29), and 98-37120 (3.0 +/- 0.41) induced significantly more severe lesions than Ingelvac PRRS MLV (0.7 +/- 0.17) and controls (0.7 +/- 0.15). The molecular analyses and biological characterizations suggest that the vaccine-like isolate 98-38803 (99.5% amino acid homology based on the ORF5 gene) induces microscopic pneumonia lesions similar in type to, but different in severity

  2. ANTIMICROBIAL PROPERTIES OF HYDROXYAPATITE COATINGS CONTAINING OF CHITOSAN AND SILVER ON TITANIUM SUBSTRATES IN RELATION TO MICROORGANISMS E.COLI ATCC 25922

    Directory of Open Access Journals (Sweden)

    Sukhodub LB

    2013-03-01

    Full Text Available In this work it was studied the antibacterial properties of coatings based on HA, with Chitosan and silver ions additions, produced by substrates termodeposition method from aqueous solutions with varying concentrations of Chitosan (0.025 and 0.1 g/l and silver (1 mg/l as the antimicrobial components as well as three-part cover, consisting of a film of Chitosan, HA and silver. Study on antibacterial properties of composite coatings on the pathogen E.coli ATCC 25922 was held by Spectrophotometric measurement and analysis of optical density of suspensions, containing samples. 3 series of measurements data were averaged. The results showed that the concentration of antimicrobial components have indicated a bacteriostatic effect of coatings on the culture of E. coli AS ATCC 25922 in physiological solution at a temperature of 37 °C. The most effective was the three-part cover consisting of a film of chitosan, HA and silver.

  3. Die another day: Fate of heat-treated Geobacillus stearothermophilus ATCC 12980 spores during storage under growth-preventing conditions.

    Science.gov (United States)

    Mtimet, Narjes; Trunet, Clément; Mathot, Anne-Gabrielle; Venaille, Laurent; Leguérinel, Ivan; Coroller, Louis; Couvert, Olivier

    2016-06-01

    Geobacillus stearothermophilus spores are recognized as one of the most wet-heat resistant among aerobic spore-forming bacteria and are responsible for 35% of canned food spoilage after incubation at 55 °C. The purpose of this study was to investigate and model the fate of heat-treated survivor spores of G. stearothermophilus ATCC 12980 in growth-preventing environment. G. stearothermophilus spores were heat-treated at four different conditions to reach one or two decimal reductions. Heat-treated spores were stored in nutrient broth at different temperatures and pH under growth-preventing conditions. Spore survival during storage was evaluated by count plating over a period of months. Results reveal that G. stearothermophilus spores surviving heat treatment lose their viability during storage under growth-preventing conditions. Two different subpopulations were observed during non-thermal inactivation. They differed according to the level of their resistance to storage stress, and the proportion of each subpopulation can be modulated by heat treatment conditions. Finally, tolerance to storage stress under growth-preventing conditions increases at refrigerated temperature and neutral pH regardless of heat treatment conditions. Such results suggest that spore inactivation due to heat treatment could be completed by storage under growth-preventing conditions. PMID:26919821

  4. Cloning, expression, purification, crystallization and preliminary X-ray characterization of allantoinase from Bacillus licheniformis ATCC 14580.

    Science.gov (United States)

    Conejero-Muriel, Mayte; Martínez-Gómez, Ana Isabel; Martínez-Rodríguez, Sergio; Gavira, Jose A

    2014-11-01

    Allantoinase, a member of the amidohydrolase superfamily, exists in a wide variety of organisms, including bacteria, fungi, plants and a few animals, such as fishes and amphibians. Allantoinase catalyzes the reversible hydrolysis of allantoin into allantoate by hydrolytic cleavage of the N1-C2 amide bond of the five-membered hydantoin ring. Allantoinase from Bacillus licheniformis (AllBali) presents an inverted enantioselectivity towards allantoin (R-enantioselective), which is a distinguishable feature that is not observed for other allantoinases. In this work, B. licheniformis ATCC 14580 allantoinase (AllBali) containing a C-terminal His6 tag was overproduced in Escherichia coli and purified to homogeneity. Crystals of AllBali were obtained by the vapour-diffusion method using 0.1 M potassium thiocyanate, 20%(w/v) polyethylene glycol 3350 as a crystallization solution. X-ray diffraction data were collected to a resolution of 3.5 Å with an Rmerge of 29.2% from a crystal belonging to space group P12₁1, with unit-cell parameters a=54.93, b=164.74, c=106.89 Å, β=98.49°. There are four molecules in the asymmetric unit with a solvent content of 47% as estimated from the Matthews coefficient (VM=2.34 Å3 Da(-1)). PMID:25372819

  5. Coupling of Cellular Processes and Their Coordinated Oscillations under Continuous Light in Cyanothece sp. ATCC 51142, a Diazotrophic Unicellular Cyanobacterium.

    Directory of Open Access Journals (Sweden)

    S Krishnakumar

    Full Text Available Unicellular diazotrophic cyanobacteria such as Cyanothece sp. ATCC 51142 (henceforth Cyanothece, temporally separate the oxygen sensitive nitrogen fixation from oxygen evolving photosynthesis not only under diurnal cycles (LD but also in continuous light (LL. However, recent reports demonstrate that the oscillations in LL occur with a shorter cycle time of ~11 h. We find that indeed, majority of the genes oscillate in LL with this cycle time. Genes that are upregulated at a particular time of day under diurnal cycle also get upregulated at an equivalent metabolic phase under LL suggesting tight coupling of various cellular events with each other and with the cell's metabolic status. A number of metabolic processes get upregulated in a coordinated fashion during the respiratory phase under LL including glycogen degradation, glycolysis, oxidative pentose phosphate pathway, and tricarboxylic acid cycle. These precede nitrogen fixation apparently to ensure sufficient energy and anoxic environment needed for the nitrogenase enzyme. Photosynthetic phase sees upregulation of photosystem II, carbonate transport, carbon concentrating mechanism, RuBisCO, glycogen synthesis and light harvesting antenna pigment biosynthesis. In Synechococcus elongates PCC 7942, a non-nitrogen fixing cyanobacteria, expression of a relatively smaller fraction of genes oscillates under LL condition with the major periodicity being 24 h. In contrast, the entire cellular machinery of Cyanothece orchestrates coordinated oscillation in anticipation of the ensuing metabolic phase in both LD and LL. These results may have important implications in understanding the timing of various cellular events and in engineering cyanobacteria for biofuel production.

  6. Purification, characterization, primary structure, crystallization and preliminary crystallographic study of a serine proteinase from Streptomyces fradiae ATCC 14544.

    Science.gov (United States)

    Kitadokoro, K; Tsuzuki, H; Nakamura, E; Sato, T; Teraoka, H

    1994-02-15

    A proteinase having wide substrate specificity was isolated from Streptomyces fradiae ATCC 14544. This proteinase, which we propose to call SFase-2, was purified from the culture filtrate by S-Sepharose chromatography. The purified enzyme showed an apparent molecular mass of 19 kDa on SDS/PAGE. When synthetic peptides were used as substrates, SFase-2 showed broad substrate specificity. It also hydrolyzed keratin, elastin and collagen as proteinaceous substrates. It was completely inhibited by diisopropylfluorophosphate and chymostatin, but not by tosylphenylalaninechloromethane, tosyllysinechloromethane or EDTA, indicating that it can be classified as a serine proteinase. The matured protein sequence of SFase-2 was determined by a combination of amino acid sequencing and the DNA sequencing of the gene. SFase-2, consisting of 191 amino acids, is a novel proteinase. It showed 76% similarity in the amino acid sequence with Streptomyces griseus proteinase A [Johnson P. and Smillie L. B. (1974) FEBS Lett. 47, 1-6]. For insight into the three-dimensional structure of SFase-2, we obtained single crystals by the vapor diffusion method using sodium phosphate as a precipitant. These crystals belonged to the orthorhombic, space group P2(1)2(1)2(1) with cell dimensions a = 6.92 nm, b = 7.28 nm, c = 2.99 nm; one molecule was present in the asymmetric unit.

  7. Modeling the behavior of Geobacillus stearothermophilus ATCC 12980 throughout its life cycle as vegetative cells or spores using growth boundaries.

    Science.gov (United States)

    Mtimet, Narjes; Trunet, Clément; Mathot, Anne-Gabrielle; Venaille, Laurent; Leguérinel, Ivan; Coroller, Louis; Couvert, Olivier

    2015-06-01

    Geobacillus stearothermophilus is recognized as one of the most prevalent micro-organism responsible for flat sour in the canned food industry. To control these highly resistant spore-forming bacteria, the heat treatment intensity could be associated with detrimental conditions for germination and outgrowth. The purpose of this work was to study successively the impact of temperature and pH on the growth rate of G. stearothermophilus ATCC 12980, its sporulation ability, its heat resistance in response to various sporulation conditions, and its recovery ability after a heat treatment. The phenotypic investigation was carried out at different temperatures and pHs on nutrient agar and the heat resistance was estimated at 115 °C. The greatest spore production and the highest heat resistances were obtained at conditions of temperature and pH allowing maximal growth rate. The current observations also revealed that growth, sporulation and recovery boundaries are close. Models using growth boundaries as main parameters were extended to describe and quantify the effect of temperature and pH throughout the life cycle of G. stearothermophilus as vegetative cells or as spore after a heat treatment and during recovery.

  8. Biotransformation of natural compounds: unexpected thio conjugation of Sch-642305 with 3-mercaptolactate catalyzed by Aspergillus niger ATCC 16404 cells.

    Science.gov (United States)

    Adelin, Emilie; Martin, Marie-Thérèse; Bricot, Marie-Françoise; Cortial, Sylvie; Retailleau, Pascal; Ouazzani, Jamal

    2012-12-01

    Sch-642305 is produced by the endophytic fungi Phomopsis sp. CMU-LMA and exhibits both antimicrobial and cytotoxic activities. The incubation of Sch-642305 with Aspergillus niger ATCC 16404 resting cells leads to two unexpected thio conjugates. Compound (1) is formed by the addition of the cysteine metabolite 3-mercaptolactate to the double bond of Sch-642305. Compound (1) undergoes an intramolecular rearrangement to give compound (2), which contains two rings: a five-membered hydroxylactone ring and a five-membered thiophene ring. The absolute configuration of compound (1) is similar to that of the parent compound, but the configuration of the mercaptolactate side-chain was not determined. The absolute configuration of compound (2) was deduced from the crystal structure and confirmed by the anomal effect of the sulfur atom. To the best of our knowledge, this is the first time such a conjugation rearrangement reactions were observed. The biological significance and the reaction mechanisms are discussed. Compound (1) exhibits a weak antimicrobial activity against Gram-positive bacteria, whereas derivatives (1) and (2) showed an IC₅₀ of 1 and 1.2 μM, respectively, against colonic epithelial cancer cells. PMID:22975164

  9. Performance analyses of a neutralizing agent combination strategy for the production of succinic acid by Actinobacillus succinogenes ATCC 55618.

    Science.gov (United States)

    Wang, Cheng-Cheng; Zhu, Li-Wen; Li, Hong-Mei; Tang, Ya-Jie

    2012-05-01

    A neutralizing agent combination strategy was developed to enhance the succinic acid production by Actinobacillus succinogenes ATCC 55618. First, a maximal succinic acid production of 48.2 g/L was obtained at a culture pH of 7.5. Second, NaOH and KOH were screened to identify the optimal neutralizing agent for pH control. However, the production of succinic acid did not increase, and severe cell flocculation was observed due to a high concentration of metal ions when only one neutralizing agent was used to control pH. Finally, a neutralizing agent combination strategy was developed with a supply of neutralizing agents with OH(-) and carbonate. The cell flocculation was eliminated, and a maximum succinic acid production of 59.2 g/L was obtained with 5 M NaOH and 40 g/L of MgCO(3); this production was 27.9% higher than that obtained with NaOH alone. The results obtained in this study may be useful for the large-scale industrial production of succinic acid. PMID:22002101

  10. Actinobacillus succinogenes ATCC 55618 fermentation medium optimization for the production of succinic acid by response surface methodology.

    Science.gov (United States)

    Zhu, Li-Wen; Wang, Cheng-Cheng; Liu, Rui-Sang; Li, Hong-Mei; Wan, Duan-Ji; Tang, Ya-Jie

    2012-01-01

    As a potential intermediary feedstock, succinic acid takes an important place in bulk chemical productions. For the first time, a method combining Plackett-Burman design (PBD), steepest ascent method (SA), and Box-Behnken design (BBD) was developed to optimize Actinobacillus succinogenes ATCC 55618 fermentation medium. First, glucose, yeast extract, and MgCO(3) were identified to be key medium components by PBD. Second, preliminary optimization was run by SA method to access the optimal region of the key medium components. Finally, the responses, that is, the production of succinic acid, were optimized simultaneously by using BBD, and the optimal concentration was located to be 84.6 g L(-1) of glucose, 14.5 g L(-1) of yeast extract, and 64.7 g L(-1) of MgCO(3). Verification experiment indicated that the maximal succinic acid production of 52.7 ± 0.8 g L(-1) was obtained under the identified optimal conditions. The result agreed with the predicted value well. Compared with that of the basic medium, the production of succinic acid and yield of succinic acid against glucose were enhanced by 67.3% and 111.1%, respectively. The results obtained in this study may be useful for the industrial commercial production of succinic acid. PMID:23093852

  11. Integrated bioconversion of pulp and paper primary sludge to second generation bioethanol using Saccharomyces cerevisiae ATCC 26602.

    Science.gov (United States)

    Mendes, Cátia V T; Cruz, Crispin H G; Reis, Diana F N; Carvalho, M Graça V S; Rocha, Jorge M S

    2016-11-01

    Primary sludge, from different pulp and paper mills, was used as feedstock in simultaneous saccharification and fermentation (SSF) processes to produce ethanol. SSF was carried out with Saccharomyces cerevisiae ATCC 26602 yeast and NS 22192 enzymatic extract using 150gL(-1) of carbohydrates (CH) from primary sludge. The effect of sterilization, reduction of enzyme dosage and fed-batch vs. batch conditions were studied. The removal of sterilization can be considered since no contamination or atypical by-products were observed, although SSF efficiency slightly decreased. The reduction of the enzyme dosage from 35 to 15FPUgCH(-1) was successful. Despite of initial mixing difficulties, batch SSF enabled higher ethanol concentration (41.7gL(-1)), conversion yield (48.9%) and productivity (0.78gL(-1)h(-1)), compared to the fed-batch process at the same conditions of low enzyme dosage of 5FPUgCH(-1) and high solids content of 21.7%, rarely found in literature.

  12. Integrated bioconversion of pulp and paper primary sludge to second generation bioethanol using Saccharomyces cerevisiae ATCC 26602.

    Science.gov (United States)

    Mendes, Cátia V T; Cruz, Crispin H G; Reis, Diana F N; Carvalho, M Graça V S; Rocha, Jorge M S

    2016-11-01

    Primary sludge, from different pulp and paper mills, was used as feedstock in simultaneous saccharification and fermentation (SSF) processes to produce ethanol. SSF was carried out with Saccharomyces cerevisiae ATCC 26602 yeast and NS 22192 enzymatic extract using 150gL(-1) of carbohydrates (CH) from primary sludge. The effect of sterilization, reduction of enzyme dosage and fed-batch vs. batch conditions were studied. The removal of sterilization can be considered since no contamination or atypical by-products were observed, although SSF efficiency slightly decreased. The reduction of the enzyme dosage from 35 to 15FPUgCH(-1) was successful. Despite of initial mixing difficulties, batch SSF enabled higher ethanol concentration (41.7gL(-1)), conversion yield (48.9%) and productivity (0.78gL(-1)h(-1)), compared to the fed-batch process at the same conditions of low enzyme dosage of 5FPUgCH(-1) and high solids content of 21.7%, rarely found in literature. PMID:27566524

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

    Science.gov (United States)

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

    2016-03-01

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

  14. EFFECT OF CULTURE MEDIUM ON BACTERIOCIN PRODUCTION BY LACTOBACILLUS RHAMNOSUS HN001 AND LACTOBACILLUS REUTERI ATCC 53608

    Directory of Open Access Journals (Sweden)

    Aguilar-Uscanga B. R.

    2013-06-01

    Full Text Available The aim of this study was to evaluate the effect of media on bacteriocin production by Lactobacillus rhamnosus HN001 and Lactobacillus reuteri ATCC 53608 using three different media: YPM, YPF and MRS supplemented with glucose and K2HPO4. The optimum temperature was 37°C and initial pH 6.5. Bacteriocin-like substances produced by tested bacteria in MRS medium supplemented with glucose and K2HPO4 exhibited a broad antimicrobial spectrum determined by well diffusion assay against indicator bacteria Listeria monocytogenes, Lactobacillus sakei, Enterococcus faecium, Lactobacillus delbrueckii, Lactobacillus acidophilus, but no antimicrobial spectrum against E. coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, Bacillus cereus was detected. Bacteriocin was sensitive to protease IV, trypsin, pepsin and -amylases, but resistant to lipase. It was also resistant to detergents such as Tween 80, Triton-X and SDS. This bacteriocin was thermo-stable (resistant at 60°C, 90°C and 100°C for 30 min. Tested bacteria showed the best antimicrobial (bacteriocin-like activity after growth in MRS medium. Bacteriocin substances produced by tested bacteria showed promising thermo-stable technological properties.

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

    Science.gov (United States)

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

    2016-03-01

    α-Amylase from Bacillus licheniformis ATCC 9945a (BliAmy) was proven to be very efficient in hydrolysis of granular starch below the temperature of gelatinization. By applying two-stage feeding strategy to achieve high-cell-density cultivation of Escherichia coli and extracellular production of BliAmy, total of 250.5 U/mL (i.e. 0.7 g/L) of enzyme was obtained. Thermostability of amylase was exploited to simplify purification. The hydrolysis of concentrated raw starch was optimized using response surface methodology. Regardless of raw starch concentration tested (20, 25, 30 %), BliAmy was very effective, achieving the final hydrolysis degree of 91 % for the hydrolysis of 30 % starch suspension after 24 h. The major A-type crystalline structure and amorphous domains of the starch granule were degraded at the same rates, while amylose-lipid complexes were not degraded. BliAmy presents interesting performances on highly concentrated solid starch and could be of value for starch-consuming industries while response surface methodology (RSM) could be efficiently applied for the optimization of the hydrolysis. PMID:26545758

  16. Influence of nutritive factors on C50 carotenoids production by Haloferax mediterranei ATCC 33500 with two-stage cultivation.

    Science.gov (United States)

    Fang, Chun-Jen; Ku, Kuo-Lung; Lee, Min-Hsiung; Su, Nan-Wei

    2010-08-01

    The production of pigments by Haloferax mediterranei ATCC 33500 with two-stage cultivation in response to nutritive factors in culture media was studied. Sodium chloride and magnesium sulfate in the second-stage media showed a marked effect upon the production of pigments, and sodium acetate could enhance the production. As the cells were harvested at mid-log phase of growth in first-stage cultivation and transferred to the defined media containing 5% sodium chloride, 0.1% sodium acetate and 8% magnesium sulfate at 37 degrees C, 120 rpm for further 24 h of cultivation, H. mediterranei exhibited to be an efficient producer of pigments. The yield of pigments could reach up to 0.604 A(494 nm) mL(-1) broth. TLC analysis and the UV-Vis spectra of individual spots thereof revealed that H. mediterranei produced three red pigments of C(50) carotenoid, namely bisanhydrobacterioruberin, monoanhydrobacterioruberin and bacterioruberin, as well as a C(45) carotenoid, 2-isopentenyl-3,4-dehydrorhodopin. PMID:20362434

  17. Desulphurization of some low-rank Turkish lignites with crude laccase produced from Trametes versicolor ATCC 200801

    Energy Technology Data Exchange (ETDEWEB)

    Aytar, Pinar; Gedikli, Serap [Graduate School of Natural and Applied Sciences, Eskisehir Osmangazi University (Turkey); Sam, Mesut [Department of Biology, Faculty of Arts and Science, Aksaray University (Turkey); Uenal, Arzu [Ministry of Agriculture and Rural Affairs, General Directorate of Agricultural Research, Ankara (Turkey); Cabuk, Ahmet [Department of Biology, Faculty of Arts and Science, Eskisehir Osmangazi University (Turkey); Kolankaya, Nazif [Department of Biology, Division of Biotechnology, Faculty of Science, Hacettepe University, Ankara (Turkey); Yueruem, Alp [Grand Water Research Institute, Technion Israel Institute of Technology, Haifa (Israel)

    2011-01-15

    In this paper, data obtained during the oxidative desulphurization of some low-rank Turkish lignites with crude laccase enzyme produced from Trametes versicolor ATCC 200801 are presented. In order to optimize desulphurization conditions, effects of incubation time, pulp density, incubation temperature, medium pH, and also lignite source on the desulphurization have been examined. The values for incubation period, pulp density, temperature and pH in optimum incubation condition were found as 30 min, 5%, 35 C, and pH 5.0, respectively. Under optimum conditions, treatment of coal samples with crude laccase has caused nearly 29% reduction in their total sulphur content. During the study, the rate of desulphurization of coal sample provided from Tuncbilek with crude laccase was found to be relatively higher than the other examined coal samples. Results of analytical assays have indicated that the treatment of coals with crude laccase has caused no change in their calorific values but reduced their sulphur emissions. 35%, 13%, and 25% reductions of pyritic sulphur, sulphate and organic sulphur in a period of 30 min were achieved, for a particle size of 200 {mu}m under optimal conditions with enzymatic desulphurization. Also, statistical analyses such as Tukey Multiple Comparison tests and ANOVA were performed. (author)

  18. Effect of low-concentration rhamnolipid on adsorption of Pseudomonas aeruginosa ATCC 9027 on hydrophilic and hydrophobic surfaces.

    Science.gov (United States)

    Zhong, Hua; Jiang, Yongbing; Zeng, Guangming; Liu, Zhifeng; Liu, Liuxia; Liu, Yang; Yang, Xin; Lai, Mingyong; He, Yibin

    2015-03-21

    The effects of low-concentration monorhamnolipid (monoRL) on the adsorption of Pseudomonas aeruginosa ATCC 9027 grown on glucose or hexadecane to glass beads with hydrophobic or hydrophilic surfaces was investigated using batch adsorption experiments. Results showed that adsorption isotherms of the cells on both types of glass beads fitted the Freundlich equation better than the Langmuir equation. The Kf of the Freundlich equation for adsorption of hexadecane-grown cell to glass beads with hydrophobic surface was remarkably higher than that for adsorption of hexadecane-grown cell to glass beads with hydrophilic surface, or glucose-grown cell to glass beads with either hydrophilic or hydrophobic surface. Furthermore, it decreased with the increasing monoRL concentration. For both groups of cells, the zeta potential was close to each other and stable with the increase of monoRL concentration. The surface hydrophobicity of hexadecane-grown cells, however, was significantly higher than that of the glucose-grown cells and it decreased with the increase of monoRL concentration. The results indicate the importance of hydrophobic interaction on adsorption of bacterial cells to surfaces and monoRL plays a role in reducing the bacterial adsorption by affecting cell surface hydrophobicity.

  19. Replacement of Soybean Meal with Animal Origin Protein Meals Improved Ramoplanin A2 Production by Actinoplanes sp. ATCC 33076.

    Science.gov (United States)

    Erkan, Deniz; Kayali, Hulya Ayar

    2016-09-01

    Ramoplanin A2 is the last resort antibiotic for treatment of many high morbidity- and mortality-rated hospital infections, and it is expected to be marketed in the forthcoming years. Therefore, high-yield production of ramoplanin A2 gains importance. In this study, meat-bone meal, poultry meal, and fish meal were used instead of soybean meal for ramoplanin A2 production by Actinoplanes sp. ATCC 33076. All animal origin nitrogen sources stimulated specific productivity. Ramoplanin A2 levels were determined as 406.805 mg L(-1) in fish meal medium and 374.218 mg L(-1) in poultry meal medium. These levels were 4.25- and 4.09-fold of basal medium, respectively. However, the total yield of poultry meal was higher than that of fish meal, which is also low-priced. In addition, the variations in pH levels, protein levels, reducing sugar levels, extracellular protease, amylase and lipase activities, and intracellular free amino acid levels were monitored during the incubation period. The correlations between ramoplanin production and these variables with respect to the incubation period were determined. The intracellular levels of L-Phe, D-Orn, and L-Leu were found critical for ramoplanin A2 production. The strategy of using animal origin nitrogen sources can be applied for large-scale ramoplanin A2 production. PMID:27142271

  20. Assessment of the CO2 fixation capacity of Anabaena sp. ATCC 33047 outdoor cultures in vertical flat-panel reactors.

    Science.gov (United States)

    Clares, Marta E; Moreno, José; Guerrero, Miguel G; García-González, Mercedes

    2014-10-10

    The extent of biological CO2 fixation was evaluated for outdoor cultures of the cyanobacterium Anabaena sp. ATCC 33047. Culture conditions were optimized indoors in bubble-column photochemostats operating in continuous mode, subjected to irradiance cycles mimicking the light regime outdoors. Highest values achieved for CO2 fixation rate and biomass productivity were 1 and 0.6 g L(-1) day(-1), respectively. The comparison among different reactors operating simultaneously - open pond, horizontal tubular reactor and vertical flat-panel - allowed to assess their relative efficiency for the outdoor development of Anabaena cultures. Despite the higher volumetric CO2 fixation capacity (and biomass productivity) exhibited by the tubular photobioreactor, yield of the flat-panel reactor was 50% higher than that of the tubular option on a per area basis, reaching values over 35 g CO2 fixed m(-2) d(-1). The flat-panel reactor actually represents a most suitable system for CO2 capture coupled to the generation of valuable biomass by Anabaena cultures.

  1. Influence of Selenium Content in the Culture Medium on Protein Profile of Yeast Cells Candida utilis ATCC 9950

    Directory of Open Access Journals (Sweden)

    Marek Kieliszek

    2015-01-01

    Full Text Available Selenium is an essential trace element for human health and it has been recognized as a component of several selenoproteins with crucial biological functions. It has been identified as a component of active centers of many enzymes, as well as integral part of biologically active complexes. The aim of the study was to evaluate the protein content and amino acid profile of the protein of fodder yeast Candida utilis ATCC 9950 cultured in media control and experimental enriched selenium. Protein analysis was performed using SDS-PAGE method consisting of polyacrylamide gel electrophoresis in the presence of SDS. The highest contents of soluble protein (49,5 mg/g were found in yeast cells after 24-hour culture conducted in control (YPD medium. In the presence of selenium there were determined small amounts of protein content. With increasing time of yeast culture (to 72 hours the control and experimental media were reported to reduce soluble protein content. In electropherogram proteins from control cultures was observed the presence of 10 protein fractions, but in all the experimental cultures (containing 20, 30, and 40 mg/L selenium of 14 protein fractions. On the basis of the molecular weights of proteins, it can be concluded that they were among others: selenoprotein 15 kDa and selenoprotein 18 kDa.

  2. Removal of antibiotic resistance gene-carrying plasmids from Lactobacillus reuteri ATCC 55730 and characterization of the resulting daughter strain, L. reuteri DSM 17938.

    Science.gov (United States)

    Rosander, Anna; Connolly, Eamonn; Roos, Stefan

    2008-10-01

    The spread of antibiotic resistance in pathogens is primarily a consequence of the indiscriminate use of antibiotics, but there is concern that food-borne lactic acid bacteria may act as reservoirs of antibiotic resistance genes when distributed in large doses to the gastrointestinal tract. Lactobacillus reuteri ATCC 55730 is a commercially available probiotic strain which has been found to harbor potentially transferable resistance genes. The aims of this study were to define the location and nature of beta-lactam, tetracycline, and lincosamide resistance determinants and, if they were found to be acquired, attempt to remove them from the strain by methods that do not genetically modify the organism before subsequently testing whether the probiotic characteristics were retained. No known beta-lactam resistance genes was found, but penicillin-binding proteins from ATCC 55730, two additional resistant strains, and three sensitive strains of L. reuteri were sequenced and comparatively analyzed. The beta-lactam resistance in ATCC 55730 is probably caused by a number of alterations in the corresponding genes and can be regarded as not transferable. The strain was found to harbor two plasmids carrying tet(W) tetracycline and lnu(A) lincosamide resistance genes, respectively. A new daughter strain, L. reuteri DSM 17938, was derived from ATCC 55730 by removal of the two plasmids, and it was shown to have lost the resistances associated with them. Direct comparison of the parent and daughter strains for a series of in vitro properties and in a human clinical trial confirmed the retained probiotic properties of the daughter strain. PMID:18689509

  3. Removal of Antibiotic Resistance Gene-Carrying Plasmids from Lactobacillus reuteri ATCC 55730 and Characterization of the Resulting Daughter Strain, L. reuteri DSM 17938▿

    OpenAIRE

    Rosander, Anna; Connolly, Eamonn; Roos, Stefan

    2008-01-01

    The spread of antibiotic resistance in pathogens is primarily a consequence of the indiscriminate use of antibiotics, but there is concern that food-borne lactic acid bacteria may act as reservoirs of antibiotic resistance genes when distributed in large doses to the gastrointestinal tract. Lactobacillus reuteri ATCC 55730 is a commercially available probiotic strain which has been found to harbor potentially transferable resistance genes. The aims of this study were to define the location an...

  4. Differentiation of Lactobacillus helveticus, Lactobacillus delbrueckii subsp bulgaricus, subsp lactis and subsp delbrueckii using physiological and genetic tools and reclassification of some strains from the ATCC collection.

    Science.gov (United States)

    Delley, Michèle; Germond, Jacques-Edouard

    2002-08-01

    Several physiological tests of glucose metabolism and genetic tools including species specific probes and 16S rDNA sequences were used to identify strains of L. helveticus and the group of L. delbrueckii with its three subspecies lactis, bulgaricus, and delbrueckii. These species are important for the milk industry as fermenting lactic acid bacteria. The identification procedure was applied to the different strains of these species available from the ATCC collection and allowed to reclassify part of them.

  5. Conclusion on the peer review of the pesticide risk assessment of the active substances Beauveria bassiana strains ATCC-74040 and GHA

    Directory of Open Access Journals (Sweden)

    European Food Safety Authority

    2013-01-01

    Full Text Available The conclusions of the European Food Safety Authority (EFSA following the peer review of the initial risk assessments carried out by the competent authority of the rapporteur Member State Germany, for the pesticide active substances Beauveria bassiana strains ATCC-74040 and GHA are reported. The context of the peer review was that required by Commission Regulation (EC No 2229/2004, as amended by Commission Regulation (EC No 1095/2007 and Commission Regulation (EU No 114/2010. The conclusions were reached on the basis of the evaluation of the representative uses of Beauveria bassiana strains ATCC-74040 and GHA as an insecticide on tomatoes for strain ATCC-74040 and on tomatoes, cucumbers and ornamentals for strain GHA. The reliable endpoints concluded as being appropriate for use in regulatory risk assessment, derived from the available studies and literature in the dossier peer reviewed, are presented. Missing information identified as being required by the regulatory framework is listed. Concerns are identified.

  6. Feasibility of installing and maintaining anaerobiosis using Escherichia coli HD701 as a facultative anaerobe for hydrogen production by Clostridium acetobutylicum ATCC 824 from various carbohydrates.

    Science.gov (United States)

    Hassan, Sedky H A; Morsy, Fatthy Mohamed

    2015-12-01

    Using Escherichia coli for installing and maintaining anaerobiosis for hydrogen production by Clostridium acetobutylicum ATCC 824 is a cost-effective approach for industrial hydrogen production, as it does not require reducing agents or sparging with inert gases. This study was devoted for investigating the feasibility for installing and maintaining anaerobiosis of hydrogen production by C. acetobutylicum ATCC 824 when using E. coli HD701 utilizable versus non utilizable sugars as a-carbon source. Using E. coli HD701 for installing anaerobiosis showed a comparable hydrogen production yield and efficiency to the use of reducing agents and nitrogen sparging in case of hydrogen production from the E. coli HD701 non utilizable sugars. In contrast, using E. coli HD701 for installing anaerobiosis showed a lower hydrogen production yield and efficiency than the use of reducing agents and nitrogen sparging in case of using glucose as a substrate. This is possibly because E. coli HD701 when using glucose compensate for the substrate, and produce hydrogen with lower efficiency than C. acetobutylicum ATCC 824. These results indicated that the use of E. coli HD701 for installing anaerobiosis would not be economically feasible when using E. coli HD701 utilizable sugars as a carbon source. In contrast, the use of this approach for installing anaerobiosis for hydrogen production from sucrose and starch would have a high potency for industrial applications.

  7. Crystal Structure of the Zorbamycin-Binding Protein ZbmA, the Primary Self-Resistance Element in Streptomyces flavoviridis ATCC21892

    Energy Technology Data Exchange (ETDEWEB)

    Rudolf, Jeffrey D. [Scripps Research Inst., Jupiter, FL (United States); Bigelow, Lance [Argonne National Lab. (ANL), Argonne, IL (United States); Chang, Changsoo [Argonne National Lab. (ANL), Argonne, IL (United States); Cuff, Marianne E. [Argonne National Lab. (ANL), Argonne, IL (United States); Lohman, Jeremy R. [Scripps Research Inst., Jupiter, FL (United States); Chang, Chin-Yuan [Scripps Research Inst., Jupiter, FL (United States); Ma, Ming [Scripps Research Inst., Jupiter, FL (United States); Yang, Dong [Scripps Research Inst., Jupiter, FL (United States); Clancy, Shonda [Argonne National Lab. (ANL), Argonne, IL (United States); Babnigg, Gyorgy [Argonne National Lab. (ANL), Argonne, IL (United States); Joachimiak, Andrzej [Argonne National Lab. (ANL), Argonne, IL (United States); Phillips, George N. [Rice Univ., Houston, TX (United States); Shen, Ben [Scripps Research Inst., Jupiter, FL (United States)

    2015-11-17

    The bleomycins (BLMs), tallysomycins (TLMs), phleomycin, and zorbamycin (ZBM) are members of the BLM family of glycopeptide-derived antitumor antibiotics. The BLM-producing Streptomyces verticillus ATCC15003 and the TLM-producing Streptoalloteichus hindustanus E465-94 ATCC31158 both possess at least two self-resistance elements, an N-acetyltransferase and a binding protein. The N-acetyltransferase provides resistance by disrupting the metal-binding domain of the antibiotic that is required for activity, while the binding protein confers resistance by sequestering the metal-bound antibiotic and preventing drug activation via molecular oxygen. We recently established that the ZBM producer, Streptomyces flavoviridis ATCC21892, lacks the N-acetyltransferase resistance gene and that the ZBM-binding protein, ZbmA, is sufficient to confer resistance in the producing strain. To investigate the resistance mechanism attributed to ZbmA, we determined the crystal structures of apo and Cu(II)-ZBM-bound ZbmA at high resolutions of 1.90 and 1.65 angstrom, respectively. A comparison and contrast with other structurally characterized members of the BLM-binding protein family revealed key differences in the protein ligand binding environment that fine-tunes the ability of ZbmA to sequester metal-bound ZBM and supports drug sequestration as the primary resistance mechanism in the producing organisms of the BLM family of antitumor antibiotics.

  8. Preliminary separation and qualitative analysis of carotenoids in Arthrobacter sp.%一株节杆菌发酵产物中类胡萝卜素的分离及组分分析的初探

    Institute of Scientific and Technical Information of China (English)

    翟玉贵; 张伟国; 钱和

    2014-01-01

    [目的]对实验室保存的一株红色球菌进行菌株鉴定,并对色素提取液的组分进行定性分析.[方法]采取形态学观察、16S rDNA序列同源性分析和生理生化分析相结合的方法确定菌株的分类地位.使用乙醇-丙酮混合液提取胞内色素,采用硅胶G薄板层析法对色素提取液进行初步分离纯化,并结合其光谱吸收特性、质谱结果进行定性分析.[结果]实验结果表明此菌株属于节杆菌属(Arthrobacter sp.).薄板层析结果显示该菌株内主要有4种色素组分,且靠近溶剂前沿组分为黄色,其它组分皆为橘红色.吸收光谱特性、质谱结果显示黄色组分可能为β-胡萝卜素,橘红色组分可能为螺菌黄质系类胡萝卜素.[结论]此株节杆菌以较为廉价的糖蜜和玉米浆作为营养物质用于满足自身生长需要,并且积累胞内类胡萝卜素,因此该菌的进一步研究有一定的价值.

  9. Pb2 + Tolerance and Adsorption of Arthrobacter sp. 12-1 Isolated from Lead-zinc Mine Tailing Dam%铅锌矿尾矿坝分离节杆菌12-1对Pb2+的耐受和吸附性能研究

    Institute of Scientific and Technical Information of China (English)

    陈志; 邹情雅; 潘晓鸿; 林璋; 关雄

    2014-01-01

    The extensive exploitation and usage of lead resources have caused serious environmental pollution problems currently. Bioremediation of Pb2 + contaminated water and soil environments using microbes is regarded as a promising technology due to the advantages of its cost-effective, easy operation and environmental-friendly properties. In order to understand Pb2+ biosorption characterization of microbes, an indigenous lead-resistant bacterium-Arthrobacter sp. 12-1(GenBank No. KM362724) was isolated from lead-zinc mine tailing dam, and the process and mechanism of Pb2+ biosorption by Arthrobacter sp. 12-1 were furtherly investigated in this study. Study on the growth of Arthrobacter sp. 12-1 in LB medium containing different concentration of Pb2+suggested that the highest Pb2+tolerant concentration of Arthrobacter sp. 12-1 was 800 mg/L. In water solution, 105 mg/L of Pb2+could be reduced to 2.17 mg/L by Arthrobacter sp. 12-1 within 24 h with biosorption rate of 97.93%. Microscopic investigation (atomic force microscopy and scanning electronic microscopy) combined with energy dispersive X-ray spectroscopy analysis showed that lead containing mineral was formed on the surface of cell after Pb2+sorption by Arthrobacter sp. 12-1. Further fourier transform infrared (FT- IR) analysis revealed that carboxyl, amide and phosphate groups of Arthrobacter sp. 12-1 might be involved in Pb2 + biosorption process. The results demonstrated that the Arthrobacter sp. 12-1 isolated from lead-zinc mine tailing dam had strong ability of Pb2 + resistance and biosorption, indicating an attractive prospect of practical applications in bioremediation Pb2+ contaminated water and soil environments. The present work provides much fundamental information for help in constructing feasible strategies for Pb2+bioremediation in the environment.%当前铅资源的粗放式开采和使用对环境造成了严重的污染。利用微生物修复铅污染具有费用低、易操作、环境友好等优

  10. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of health claims related to Lactobacillus rhamnosus GR-1 (ATCC 55826) in combination with Lactobacillus reuteri RC-14 (ATCC 55845) and defence against vaginal pathogens by increasing the proportion of lactobacilli and/or decreasing the proportion of potentially pathogenic bacteria and/or yeasts (ID 945) pursuant to Article 13(1) of Regulation (EC) No 1924/2006

    OpenAIRE

    Tetens, Inge

    2011-01-01

    Following a request from the European Commission, the Panel on Dietetic Products, Nutrition and Allergies was asked to provide a scientific opinion on a list of health claims pursuant to Article 13 of Regulation (EC) No 1924/2006. This opinion addresses the scientific substantiation of health claims in relation to Lactobacillus rhamnosus GR-1 (ATCC 55826) in combination with Lactobacillus reuteri RC-14 (ATCC 55845) and defence against vaginal pathogens by increasing the proportion of lactobac...

  11. Increased mannitol production in Lactobacillus reuteri ATCC 55730 production strain with a modified 6-phosphofructo-1-kinase.

    Science.gov (United States)

    Papagianni, Maria; Legiša, Matic

    2014-07-10

    Based on established knowledge of the simultaneous use of the phosphoketolase pathway (PKP) and the Embden-Meyerhof pathway (EMP) - as a secondary pathway with a smaller flux - by mannitol producer Lactobacillus reuteri ATCC 55730, we demonstrated the hypothesis that by enhancing the flux through the EMP the ability of the microorganism to handle elevated glucose concentrations will be improved, in addition to its growth rate and biomass yield. NADH availability will be increased and its demand will be satisfied, allowing the electron acceptor fructose to be more efficiently transformed into mannitol. A truncated version of the gene encoding 6-phospho-1-fructokinase (tpfkA) from the NRRL 2270 strain of Aspergillus niger along with its activator pkaC were introduced into the microorganism by plasmid transformation. Growth of the transformants at elevated glucose concentrations in the presence of fructose resulted in improved assimilation of the provided carbohydrates and a significant increase in the overall fermentation productivities. At the highest tested levels of glucose and fructose (75g/l each), the transformant strain experienced a 4-fold increase in PFK activity and a 2.3-fold increase in the glycolytic flux while the biomass yield reached 7g/l (1.6g/l in the parental strain), the mannitol yield was 56g/l (10g/l in the parental strain) and the lactate yield was 21g/l (3.5g/l in the parental strain). A high NADH/NAD(+) ratio occurred under increased glycolytic flux conditions and facilitated the efficient conversion of fructose to mannitol. A direct effect of deregulated PFK activity on the glycolytic flux is therefore demonstrated in the present case suggesting an alternative approach of metabolic engineering in L. reuteri for increased mannitol production. PMID:24742994

  12. A study of the surface charging properties of a standard strain of Escherichia coli (ATCC 11775) in aqueous solutions.

    Science.gov (United States)

    Li, Xinpeng; Xue, Xinkai; Pashley, Richard M

    2015-11-01

    In this study, the Escherichia coli (E. coli) strain ATCC 11775 was studied to determine its surface charging properties in a range of different aqueous salt solutions, with the aim of evaluating its potential as a monitor organism for water treatment. Zeta potential measurements were carried out in various solutions containing: NaCl, CaCl2, MgSO4, ZnSO4 and C14TAB, at different pH values and concentrations. Interestingly, it was found that the zeta potential of this strain of E. coli remained fairly constant at pH values over about 6, in 1mM NaCl solutions. In order to explain the cell surface charging properties, a simple, mass action surface ionization model was developed. This model indicates that the surface charging of these E. coli cells can be modeled simply using the ionization behavior of the acid groups in the common anionic membrane lipid phosphatidylserine (PS). There appeared to be no specific, strong adsorption of either divalent anions or cations, until high salt concentrations, above about 0.1M. The results suggest that at high concentrations both Ca(2+) and SO4(2-) ions are strongly adsorbed at the cell surface. However reduction of the magnitude of the surface electrostatic potential, due to Ca(2+) ion adsorption, did not appear to cause any cellular binding. In comparison, cationic surfactant was strongly adsorbed by the cell membrane surface, even at concentrations of 0.1mM, and light scattering studies indicated that the adsorption of the surfactant appeared to lyse the cell membrane and release internal cellular materials leading to a significant reduction in cell size.

  13. Rhythmic and sustained oscillations in metabolism and gene expression of Cyanothece sp. ATCC 51142 under constant light

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    Sandeep Bhupendra Gaudana

    2013-12-01

    Full Text Available Cyanobacteria, a group of photosynthetic prokaryotes, oscillate between day and night time metabolisms with concomitant oscillations in gene expression in response to light/dark cycles (LD. The oscillations in gene expression have been shown to sustain in constant light (LL with a free running period of 24 h in a model cyanobacterium Synechococcus elongatus PCC 7942. However, equivalent oscillations in metabolism are not reported under LL in this non-nitrogen fixing cyanobacterium. Here we focus on Cyanothece sp. ATCC 51142, a unicellular, nitrogen-fixing cyanobacterium known to temporally separate the processes of oxygenic photosynthesis and oxygen-sensitive nitrogen fixation. In a recent report, metabolism of Cyanothece 51142 has been shown to oscillate between photosynthetic and respiratory phases under LL with free running periods that are temperature dependent but significantly shorter than the circadian period. Further, the oscillations shift to circadian pattern at moderate cell densities that are concomitant with slower growth rates. Here we take this understanding forward and demonstrate that the utradian rhythm under LL sustains at much higher cell densities when grown under turbulent regimes that simulate flashing light effect. Our results suggest that the ultradian rhythm in metabolism may be needed to support higher carbon and nitrogen requirements of rapidly growing cells under LL. With a comprehensive Real time PCR based gene expression analysis we account for key regulatory interactions and demonstrate the interplay between clock genes and the genes of key metabolic pathways. Further, we observe that several genes that peak at dusk in Synechococcus peak at dawn in Cyanothece and vice versa. The circadian rhythm of this organism appears to be more robust with peaking of genes in anticipation of the ensuing photosynthetic and respiratory metabolic phases.

  14. Growth inhibitory response and ultrastructural modification of oral-associated candidal reference strains (ATCC) by Piper betle L. extract

    Institute of Scientific and Technical Information of China (English)

    Mohd-Al-Faisal Nordin; Wan Himratul-Aznita Wan Harun; Fathilah Abdul Razak; Md Yusoff Musa

    2014-01-01

    Candida species have been associated with the emergence of strains resistant to selected antifungal agents. Plant products have been used traditionally as alternative medicine to ease mucosal fungal infections. This study aimed to investigate the effects of Piper betle extract on the growth profile and the ultrastructure of commonly isolated oral candidal cells. The major component of P. betle was identified using liquid chromatography-mass spectrophotometry (LC-MS/MS). Seven ATCC control strains of Candida species were cultured in yeast peptone dextrose broth under four different growth environments:(i) in the absence of P. betle extract;and in the presence of P. betle extract at respective concentrations of (ii) 1 mg?mL21;(iii) 3 mg?mL21;and (iv) 6 mg?mL21. The growth inhibitory responses of the candidal cells were determined based on changes in the specific growth rates (m). Scanning electron microscopy (SEM) was used to observe any ultrastructural alterations in the candida colonies. LC-MS/MS was performed to validate the presence of bioactive compounds in the extract. Following treatment, it was observed that the m-values of the treated cells were significantly different than those of the untreated cells (P,0.05), indicating the fungistatic properties of the P. betle extract. The candidal population was also reduced from an average of 13.443106 to 1.783106 viable cell counts (CFU)?mL21. SEM examination exhibited physical damage and considerable morphological alterations of the treated cells. The compound profile from LC-MS/MS indicated the presence of hydroxybenzoic acid, chavibetol and hydroxychavicol in P. betle extract. The effects of P. betle on candida cells could potentiate its antifungal activity.

  15. Radiosensibilisation of bacteria on beef minced by essential oils with special reference to the spores of Bacillus cereus ATCC 7004

    International Nuclear Information System (INIS)

    The radiosensitization of Bacillus Cereus ATCC 7004 spores was evaluated in the presence of thymol, thyme, D-L menthol, trans-cinnamaldehyde and eugenol in ground beef. Meat cattle minced (5 % fat) was inoculated with spores of Bacillus Cereus (10 5 - 10 6 CFU / g), and each compound was added separately at various concentrations. The antimicrobial potential was evaluated in unirradiated meat by determining the MIC in percentage (wt / wt) after 24 h of storage at 4± 1C. Results showed that the best antimicrobial compound was the trans-cinnamaldehyde with MIC of 1.47%, wt/wt. In presence of cinnamaldehyde, the addition of sodium pyrophosphate decahydrate (0.1%, wt/wt) increased significantly (p < 0.05) the relative sensitivity of Bacillus Cereus spores 2 times. However, the presence of ascorbic acid in the media reduced significantly (p < 0.05) the radiosensitivity of bacteria. The combined effect of gamma irradiation in presence of cinnamaldehyde, added with ascorbic acid or sodium pyrophosphate decahydrate, on the microbiological and physico-chemical characteristic of meat samples was evaluated at 2 kGy under air. The use of the active compounds with the irradiation reduced significantly (p < 0.05) the count of total bacteria with a concomitant effect in the extension periods of shelf life. The addition of the cinnamaldehyde induced a significant reduction (p < 0.05) in TVN and free amino acids of irradiated samples. In presence of ascorbic acid the thiobarbituric acid-reactive substances (TBARS) concentration was significantly reduced (P...0.05). A significant reduction (p < 0.05) of a* and C* of color values and a significant increase (p < 0.05 ) of b* value were obtained for the samples treated by the cinnamaldehyde. The application of bioactive films for the immobilization of the essential oils is a good alternate to check their stability during storage time. (Author). 155 refs

  16. Chitinase production by Bacillus subtilis ATCC 11774 and its effect on biocontrol of Rhizoctonia diseases of potato.

    Science.gov (United States)

    Saber, Wesam I A; Ghoneem, Khalid M; Al-Askar, Abdulaziz A; Rashad, Younes M; Ali, Abeer A; Rashad, Ehsan M

    2015-12-01

    Stem canker and black scurf of potato, caused by Rhizoctonia solani, can be serious diseases causing an economically significant damage. Biocontrol activity of Bacillus subtilis ATCC 11774 against the Rhizoctonia diseases of potato was investigated in this study. Chitinase enzyme was optimally produced by B. subtilis under batch fermentation conditions similar to those of the potato-growing soil. The maximum chitinase was obtained at initial pH 8 and 30 °C. In vitro, the lytic action of the B. subtilis chitinase was detected releasing 355 μg GlcNAc ml⁻¹ from the cell wall extract of R. solani and suggesting the presence of various chitinase enzymes in the bacterial filtrate. In dual culture test, the antagonistic behavior of B. subtilis resulted in the inhibition of the radial growth of R. solani by 48.1% after 4 days. Moreover, the extracted B. subtilis chitinase reduced the growth of R. solani by 42.3% when incorporated with the PDA plates. Under greenhouse conditions, application of a bacterial suspension of B. subtilis at 109 cell mL⁻¹ significantly reduced the disease incidence of stem canker and black scurf to 22.3 and 30%, respectively. In addition, it significantly improved some biochemical parameters, growth and tubers yield. Our findings indicate two points; firstly, B. subtilis possesses a good biocontrol activity against Rhizoctonia diseases of potato, secondly, the harmonization and suitability of the soil conditions to the growth and activity of B. subtilis guaranteed a high controlling capacity against the target pathogen. PMID:26616375

  17. Protoplast fusion technology for improved production of coenzyme Q10 using Paracoccus denitrificans ATCC 19367 mutant strains

    Directory of Open Access Journals (Sweden)

    Pradipta Tokdar

    2014-01-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Induced mutants generated from Paracoccus denitrificans ATCC 19367 having antibiotic resistant markers, were used as parent strains to carry out protoplast fusion. The generated fusants were screened using standardized protocol for CoQ10 production. Among the generated fusants, one fusant namely PF-P1 showed 1.73 folds enhancements in specific CoQ10 content than wild type strain. Fusant PF-P1 was characterized by biochemical and molecular approaches where it showed differences than wild type strain. The fusant was further identified by 16S rRNA gene sequence analysis that showed eight nucleotide base pair mutation on conserved region and 99% homology with Paracoccus denitrificans strains. /* 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;}

  18. A quantitative metabolomics study of high sodium response in Clostridium acetobutylicum ATCC 824 acetone-butanol-ethanol (ABE) fermentation.

    Science.gov (United States)

    Zhao, Xinhe; Condruz, Stefan; Chen, Jingkui; Jolicoeur, Mario

    2016-01-01

    Hemicellulose hydrolysates, sugar-rich feedstocks used in biobutanol refinery, are normally obtained by adding sodium hydroxide in the hydrolyze process. However, the resulting high sodium concentration in the hydrolysate inhibits ABE (acetone-butanol-ethanol) fermentation, and thus limits the use of these low-cost feedstocks. We have thus studied the effect of high sodium on the metabolic behavior of Clostridium acetobutyricum ATCC 824, with xylose as the carbon source. At a threshold sodium concentration of 200 mM, a decrease of the maximum cell dry weight (-19.50 ± 0.85%) and of ABE yield (-35.14 ± 3.50% acetone, -33.37 ± 0.74% butanol, -22.95 ± 1.81% ethanol) were observed compared to control culture. However, solvents specific productivities were not affected by supplementing sodium. The main effects of high sodium on cell metabolism were observed in acidogenesis, during which we observed the accumulation of ATP and NADH, and the inhibition of the pentose phosphate (PPP) and the glycolytic pathways with up to 80.73 ± 1.47% and 68.84 ± 3.42% decrease of the associated metabolic intermediates, respectively. However, the NADP(+)-to-NADPH ratio was constant for the whole culture duration, a phenomenon explaining the robustness of solvents specific productivities. Therefore, high sodium, which inhibited biomass growth through coordinated metabolic effects, interestingly triggered cell robustness on solvents specific productivity. PMID:27321153

  19. Transcriptional analysis of the unicellular, diazotrophic cyanobacterium Cyanothece sp. ATCC 51142 grown under short day/night cycles

    Energy Technology Data Exchange (ETDEWEB)

    Toepel, Jorg; McDermott, Jason E.; Summerfield, Tina; Sherman, Louis A.

    2009-06-01

    Cyanothece sp. strain ATCC 51142 is a unicellular, diazotrophic cyanobacterium that demonstrates extensive metabolic periodicities of photosynthesis, respiration and nitrogen fixation when grown under N2-fixing conditions. We have performed a global transcription analysis of this organism using 6 h light/dark cycles in order to determine the response of the cell to these conditions and to differentiate between diurnal and circadian regulated genes. In addition, we used a context-likelihood of relatedness (CLR) analysis with this data and those from two-day light/dark and light-dark plus continuous light experiments to better differentiate between diurnal and circadian regulated genes. Cyanothece sp. adapted in several ways to growth under short light/dark conditions. Nitrogen was fixed in every second dark period and only once in each 24 h period. Nitrogen fixation was strongly correlated to the energy status of the cells and glycogen breakdown and high respiration rates were necessary to provide appropriate energy and anoxic conditions for this process. We conclude that glycogen breakdown is a key regulatory step within these complex processes. Our results demonstrated that the main metabolic genes involved in photosynthesis, respiration, nitrogen fixation and central carbohydrate metabolism have strong (or total) circadian-regulated components. The short light/dark cycles enable us to identify transcriptional differences among the family of psbA genes, as well as the differing patterns of the hup genes, which follow the same pattern as nitrogenase genes, relative to the hox genes which displayed a diurnal, dark-dependent gene expression.

  20. Role of Fimbriae, Flagella and Cellulose on the Attachment of Salmonella Typhimurium ATCC 14028 to Plant Cell Wall Models.

    Directory of Open Access Journals (Sweden)

    Michelle S F Tan

    Full Text Available Cases of foodborne disease caused by Salmonella are frequently associated with the consumption of minimally processed produce. Bacterial cell surface components are known to be important for the attachment of bacterial pathogens to fresh produce. The role of these extracellular structures in Salmonella attachment to plant cell walls has not been investigated in detail. We investigated the role of flagella, fimbriae and cellulose on the attachment of Salmonella Typhimurium ATCC 14028 and a range of isogenic deletion mutants (ΔfliC fljB, ΔbcsA, ΔcsgA, ΔcsgA bcsA and ΔcsgD to bacterial cellulose (BC-based plant cell wall models [BC-Pectin (BCP, BC-Xyloglucan (BCX and BC-Pectin-Xyloglucan (BCPX] after growth at different temperatures (28°C and 37°C. We found that all three cell surface components were produced at 28°C but only the flagella was produced at 37°C. Flagella appeared to be most important for attachment (reduction of up to 1.5 log CFU/cm2 although both cellulose and fimbriae also aided in attachment. The csgD deletion mutant, which lacks both cellulose and fimbriae, showed significantly higher attachment as compared to wild type cells at 37°C. This may be due to the increased expression of flagella-related genes which are also indirectly regulated by the csgD gene. Our study suggests that bacterial attachment to plant cell walls is a complex process involving many factors. Although flagella, cellulose and fimbriae all aid in attachment, these structures are not the only mechanism as no strain was completely defective in its attachment.

  1. Estudios hematológicos y patológicos comparativos de cerdos inoculados con un aislado de campo y el serotipo 5 ATCC de Actinobacillus pleuropneumoniae Comparative hematological and pathological study of inoculated pigs with a field isolate and an ATCC serotype 5 of Actinobacillus pleuropneumoniae

    Directory of Open Access Journals (Sweden)

    D Muñoz

    2010-01-01

    Full Text Available Se realizó una inoculación experimental de A. pleuropneumoniae utilizando un aislado de campo y una cepa de referencia ATCC serotipo 5, para lo cual se utilizaron tres grupos de animales (n = 15 para cada grupo. El grupo 1 (G1 fue inoculado con medio estéril, el grupo (G2 con serotipo 5 ATCC y el grupo 3 (G3 fue inoculado con un aislado de campo (418/07. Los resultados mostraron diferencias significativas (P ≤ 0,05 en el recuento de leucocitos totales entre el grupo G1 v/s G2 y G1 v/s G3 y los grados de las lesiones pulmonares totales evidenciaron diferencias estadísticamente significativas (P ≤ 0,05 entre los tres grupos de estudio. Las lesiones histopatológicas pulmonares mostraron diferencias estadísticas relevantes sólo entre G1 y G3 (P ≤ 0,05. En este trabajo se verifican diferencias importantes del comportamiento entre el aislado de campo y el serotipo 5 ATCC, sobre los cambios hematológicos y las lesiones macroscópicas e histopatológicas ocasionadas por ellos, lo cual podría indicar una mayor virulencia y patogenicidad del aislado nacional. Se espera en un futuro próximo serotipificar este aislado nacional de App.An experimental inoculation of Actinobacillus pleuropneumoniae (App was carried out with a field isolate and an ATCC serotype 5. Three groups of 15 pigs each were used. Group 1 (G1 was the control group inoculated with sterile media, Group 2 was inoculated with the serotype 5 ATCC, and Group 3 (G3 was inoculated with a field isolate (418/07. The results showed statistically significant differences (P ≤ 0.05 in the total leukocytes count between G1 v/s G2 and G1 v/s G3. The total macroscopic lung lesions scores were statistically different among the 3 groups (P ≤ 0.05. However, statistical difference was found only between G1 and G3 in the histopathological lung lesions (P ≤ 0.05. This work shows a clear difference in the hematological changes and the macroscopic and histopathological lesions between the

  2. ANTIBACTERIAL ACTIVITY OF AQUEOUS AND ETHANOLIC EXTRACTS OF GARLIC, CINNAMON AND TURMERIC AGAINST ESCHERICHIA COLI ATCC 25922 AND BACILLUS SUBTILIS DSM 3256

    Directory of Open Access Journals (Sweden)

    Sana Mukhtar

    2012-05-01

    Full Text Available Many of the spices used daily have been documented to be antimicrobial and have medicinal value as well. Most bacteria are sensitive to the extracts from plants such as clove, garlic, mustard, onion, oregano, turmeric etc. spices such as garlic turmeric and cinnamon has been used as antimicrobial agents in their raw form for the treatment of wounds and injuries and joint pains etc. The present study was conducted to investigate the antibacterial activity of garlic, cinnamon and turmeric. Different concentrations of extracts were prepared by using two solvents water and ethanol. The antibacterial activity was tested against Bacillus subtilus (DSM 3256 and E.coli (ATCC 25922 at different concentration of extracts of spices by using disc diffusion method. According to the results among the selected spices garlic had the best inhibitory activity showing maximum zone of 26mm against Bacillus subtilis DSM and a zone of 22mm against E.coli ATCC 25922. The aqueous extracts of garlic were more effective than ethanolic extract. In the case of cinnamon and turmeric, the ethanolic extracts were more effective exhibiting zones of 16mm against B.subtilis DSM 3256 and 17mm against E.coli , which showed that the cinnamon ethanolic extracts are equally effective against both Gram negative and Gram positive bacteria. The widest zones formed by ethanolic extract of turmeric against B.subtilis was measured as 14mm and it was 11mm for E.coli ATCC 25922. The results showed that B.subtilus is more susceptible to test spices as compared to E.coli.

  3. Efecto del sistema glucosa oxidasa/glucosa sobre el crecimiento de Escherichia coli ATCC 25922 en leche

    Directory of Open Access Journals (Sweden)

    Nirza Noguera

    2014-06-01

    Full Text Available La leche es uno de los alimentos de mayor importancia por ser rico en nutrientes y porque constituye la materia prima para la elaboración de una amplia gama de productos. Se ha demostrado que en leches pasteurizadas de marcas comerciales, pueden ocurrir contaminaciones postproceso, lo que representa un riesgo para la salud pública. Es por ello que en las últimas décadas, ha ganado importancia el uso de aditivos sintéticos u orgánicos como técnica complementaria durante el procesamiento de alimentos. La enzima glucosa oxidasa (GOX tiene amplio uso en la industria de alimentos gracias a sus propiedades antioxidante y antimicrobiana. Adicionalmente, se ha demostrado su capacidad de inhibir el crecimiento de diferentes enterobacterias. Por tal motivo, en el presente trabajo se planteó adicionar la enzima GOX en la leche y evaluar su efecto sobre el crecimiento de una cepa deEscherichia coli ATCC 25922. Se estandarizó la concentración de GOX y glucosa que ocasionaba la inhibición del crecimiento bacteriano en medio Luria-Bertani y en función de los resultados, se decidió utilizar la combinación de 2 U de GOX y 2,0 % de glucosa para agregarlos como aditivos en la leche y se establecieron dos sistemas: GOX/G sin pasteurizar y GOX/G pasteurizado. El crecimiento fue monitoreado por la técnica de contaje de colonias en placa-agar, a partir de 1 mL de cultivo a las 4, 6 y 24 h de incubación. Se encontró que los sistemas con GOX hasta las 6 horas presentaron efectos similares, inhibiendo significativamente el crecimiento de la bacteria, mientras que a las 24 h ya no se observó dicha inhibición, pero sí que el sistema con GOX pasteurizada exhibió una población menor que el sistema GOX sin pasteurizar. Estos hallazgos proyectan a la enzima GOX como una alternativa para la conservación de la leche, tanto cruda como pasteurizada.

  4. Efecto del sistema glucosa oxidasa/glucosa sobre el crecimiento de Escherichia coli ATCC 25922 en leche

    Directory of Open Access Journals (Sweden)

    Nirza Noguera

    2014-07-01

    Full Text Available La leche es uno de los alimentos de mayor importancia por ser rico en nutrientes y porque constituye la materia prima para la elaboración de una amplia gama de productos. Se ha demostrado que en leches pasteurizadas de marcas comerciales, pueden ocurrir contaminaciones postproceso, lo que representa un riesgo para la salud pública. Es por ello que en las últimas décadas, ha ganado importancia el uso de aditivos sintéticos u orgánicos como técnica complementaria durante el procesamiento de alimentos. La enzima glucosa oxidasa (GOX tiene amplio uso en la industria de alimentos gracias a sus propiedades antioxidante y antimicrobiana. Adicionalmente, se ha demostrado su capacidad de inhibir el crecimiento de diferentes enterobacterias. Por tal motivo, en el presente trabajo se planteó adicionar la enzima GOX en la leche y evaluar su efecto sobre el crecimiento de una cepa de Escherichia coli ATCC 25922. Se estandarizó la concentración de GOX y glucosa que ocasionaba la inhibición del crecimiento bacteriano en medio Luria-Bertani y en función de los resultados, se decidió utilizar la combinación de 2 U de GOX y 2,0 % de glucosa para agregarlos como aditivos en la leche y se establecieron dos sistemas: GOX/G sin pasteurizar y GOX/G pasteurizado. El crecimiento fue monitoreado por la técnica de contaje de colonias en placa-agar, a partir de 1 mL de cultivo a las 4, 6 y 24 h de incubación. Se encontró que los sistemas con GOX hasta las 6 horas presentaron efectos similares, inhibiendo significativamente el crecimiento de la bacteria, mientras que a las 24 h ya no se observó dicha inhibición, pero sí que el sistema con GOX pasteurizada exhibió una población menor que el sistema GOX sin pasteurizar. Estos hallazgos proyectan a la enzima GOX como una alternativa para la conservación de la leche, tanto cruda como pasteurizada.

  5. Transcriptional response of Corynebacterium glutamicum ATCC 13032 to hydrogen peroxide stress and characterization of the OxyR regulon.

    Science.gov (United States)

    Milse, Johanna; Petri, Kathrin; Rückert, Christian; Kalinowski, Jörn

    2014-11-20

    The aerobic soil bacterium Corynebacterium glutamicum ATCC 13032 has a remarkable natural resistance to hydrogen peroxide. A major player in hydrogen peroxide defense is the LysR type transcriptional regulator OxyR, homologs of which are present in a wide range of bacteria. In this study, the global transcriptional response of C. glutamicum to oxidative stress induced by hydrogen peroxide was examined using whole genome DNA microarrays, demonstrating the dynamic reaction of the regulatory networks. Deletion of oxyR resulted in an increased resistance of the C. glutamicum mutant to hydrogen peroxide. By performing DNA microarray hybridizations and RT-qPCR, differentially expressed genes were detected in the mutant. The direct control by OxyR was verified by electrophoretic mobility shift assays for 12 target regions. The results demonstrated that OxyR in C. glutamicum acts as a transcriptional repressor under non-stress conditions for a total of 23 genes. The regulated genes encode proteins related to oxidative stress response (e.g. katA), iron homeostasis (e.g. dps) and sulfur metabolism (e.g. suf cluster). Besides the regulator of the suf cluster, SufR, OxyR regulated the gene cg1695 encoding a putative transcriptional regulator, indicating the role of OxyR as a master regulator in defense against oxidative stress. Using a modified DNase footprint approach, the OxyR-binding sites in five target promoter regions, katA, cydA, hemH, dps and cg1292, were localized and in each upstream region at least two overlapping binding sites were found. The DNA regions protected by the OxyR protein are about 56bp in length and do not have evident sequence similarities. Still, by giving an insight in the H2O2 stimulon and extending the OxyR regulon this study considerably contributes to the understanding of the response of C. glutamicum to hydrogen peroxide-mediated oxidative stress. PMID:25107507

  6. A comparative genomics perspective on the genetic content of the alkaliphilic haloarchaeon Natrialba magadii ATCC 43099T

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    Siddaramappa Shivakumara

    2012-05-01

    Full Text Available Abstract Background Natrialba magadii is an aerobic chemoorganotrophic member of the Euryarchaeota and is a dual extremophile requiring alkaline conditions and hypersalinity for optimal growth. The genome sequence of Nab. magadii type strain ATCC 43099 was deciphered to obtain a comprehensive insight into the genetic content of this haloarchaeon and to understand the basis of some of the cellular functions necessary for its survival. Results The genome of Nab. magadii consists of four replicons with a total sequence of 4,443,643 bp and encodes 4,212 putative proteins, some of which contain peptide repeats of various lengths. Comparative genome analyses facilitated the identification of genes encoding putative proteins involved in adaptation to hypersalinity, stress response, glycosylation, and polysaccharide biosynthesis. A proton-driven ATP synthase and a variety of putative cytochromes and other proteins supporting aerobic respiration and electron transfer were encoded by one or more of Nab. magadii replicons. The genome encodes a number of putative proteases/peptidases as well as protein secretion functions. Genes encoding putative transcriptional regulators, basal transcription factors, signal perception/transduction proteins, and chemotaxis/phototaxis proteins were abundant in the genome. Pathways for the biosynthesis of thiamine, riboflavin, heme, cobalamin, coenzyme F420 and other essential co-factors were deduced by in depth sequence analyses. However, approximately 36% of Nab. magadii protein coding genes could not be assigned a function based on Blast analysis and have been annotated as encoding hypothetical or conserved hypothetical proteins. Furthermore, despite extensive comparative genomic analyses, genes necessary for survival in alkaline conditions could not be identified in Nab. magadii. Conclusions Based on genomic analyses, Nab. magadii is predicted to be metabolically versatile and it could use different carbon and energy

  7. The genome of the insecticidal Chromobacterium subtsugae PRAA4-1 and its comparison with that of Chromobacterium violaceum ATCC 12472.

    Science.gov (United States)

    Blackburn, Michael B; Sparks, Michael E; Gundersen-Rindal, Dawn E

    2016-12-01

    The genome of Chromobacterium subtsugae strain PRAA4-1, a betaproteobacterium producing insecticidal compounds, was sequenced and compared with the genome of C. violaceum ATCC 12472. The genome of C. subtsugae displayed a reduction in genes devoted to capsular and extracellular polysaccharide, possessed no genes encoding nitrate reductases, and exhibited many more phage-related sequences than were observed for C. violaceum. The genomes of both species possess a number of gene clusters predicted to encode biosynthetic complexes for secondary metabolites; these clusters suggest they produce overlapping, but distinct assortments of metabolites. PMID:27617206

  8. Expression of cbsA Encoding the Collagen-Binding S-Protein of Lactobacillus crispatus JCM5810 in Lactobacillus casei ATCC 393T

    OpenAIRE

    Martínez, Beatriz; Sillanpää, Jouko; Smit, Egbert,; Korhonen, Timo K.; Pouwels, Peter H.

    2000-01-01

    The cbsA gene encoding the collagen-binding S-layer protein of Lactobacillus crispatus JCM5810 was expressed in L. casei ATCC 393T. The S-protein was not retained on the surface of the recombinant bacteria but was secreted into the medium. By translational fusion of CbsA to the cell wall sorting signal of the proteinase, PrtP, of L. casei, CbsA was presented at the surface, rendering the transformants able to bind to immobilized collagens.

  9. Characterization of the Extended-Spectrum β-Lactamase Reference Strain, Klebsiella pneumoniae K6 (ATCC 700603), Which Produces the Novel Enzyme SHV-18

    OpenAIRE

    Rasheed, J. Kamile; Anderson, Gregory J.; Yigit, Hesna; Queenan, Anne Marie; Doménech-Sánchez, Antonio; Swenson, Jana M.; Biddle, James W.; Ferraro, Mary Jane; Jacoby, George A.; Tenover, Fred C.

    2000-01-01

    Klebsiella pneumoniae K6 (ATCC 700603), a clinical isolate, is resistant to ceftazidime and other oxyimino-β-lactams. A consistent reduction in the MICs of oxyimino-β-lactams by at least 3 twofold dilutions in the presence of clavulanic acid confirmed the utility of K. pneumoniae K6 as a quality control strain for extended-spectrum β-lactamase (ESBL) detection. Isoelectric-focusing analysis of crude lysates of K6 demonstrated a single β-lactamase with a pI of 7.8 and a substrate profile showi...

  10. Enantioselective microbial reduction of 1,1-dimethyl-1-sila-cyclohexan-2-one with growing cells of the yeast Kloeckera corticis (ATCC 20109)

    OpenAIRE

    Tacke, Reinhold; Hengelsberg, H.; Zilch, H.; Stumpf, B

    2012-01-01

    (R)-1,1-Dimethyl-1-sila-cyclohexan-2-ol [(R)-2] was prepared by enantioselective microbial reduction of 1,1-dimethyl-1-sila-cyclohexan-2-one (1) with growing cells of the yeast Kloeckera corticis (ATCC 20109). At a substrate concentration of 0.5 g/1 (temperature 27° C, incubation time 16 h), (R}-2 was obtained on a preparative scale in 60% yield and with an enantiomeric purity of 92% ee. Repeated recrystallization of the biotransformation product from n-hexane raised the enantiomeric purity t...

  11. Expression of Lactobacillus reuteri Pg4 collagen-binding protein gene in Lactobacillus casei ATCC 393 increases its adhesion ability to Caco-2 cells.

    Science.gov (United States)

    Hsueh, Hsiang-Yun; Yueh, Pei-Ying; Yu, Bi; Zhao, Xin; Liu, Je-Ruei

    2010-12-01

    The collagen-binding protein gene cnb was cloned from the probiotic Lactobacillus reuteri strain Pg4. The DNA sequence of the cnb gene (792 bp) has an open reading frame encoding 263 amino acids with a calculated molecular weight of 28.5 kDa. The cnb gene was constructed so as to constitutively express under the control of the Lactococcus lactis lacA promoter and was transformed into Lactobacillus casei ATCC 393, a strain isolated from dairy products with poor ability to adhere to intestinal epithelial cells. Confocal immunofluorescence microscopic and flow cytometric analysis of the transformed strain Lb. casei pNZ-cnb indicated that Cnb was displayed on its cell surface. Lb. casei pNZ-cnb not only showed a higher ability to adhere to Caco-2 cells but also exhibited a higher competition ability against Escherichia coli O157:H7 and Listeria monocytogenes adhesion to Caco-2 cells than Lb. casei ATCC 393. PMID:21070005

  12. Genome-scale modeling of light-driven reductant partitioning and carbon fluxes in diazotrophic unicellular cyanobacterium Cyanothece sp. ATCC 51142.

    Science.gov (United States)

    Vu, Trang T; Stolyar, Sergey M; Pinchuk, Grigoriy E; Hill, Eric A; Kucek, Leo A; Brown, Roslyn N; Lipton, Mary S; Osterman, Andrei; Fredrickson, Jim K; Konopka, Allan E; Beliaev, Alexander S; Reed, Jennifer L

    2012-01-01

    Genome-scale metabolic models have proven useful for answering fundamental questions about metabolic capabilities of a variety of microorganisms, as well as informing their metabolic engineering. However, only a few models are available for oxygenic photosynthetic microorganisms, particularly in cyanobacteria in which photosynthetic and respiratory electron transport chains (ETC) share components. We addressed the complexity of cyanobacterial ETC by developing a genome-scale model for the diazotrophic cyanobacterium, Cyanothece sp. ATCC 51142. The resulting metabolic reconstruction, iCce806, consists of 806 genes associated with 667 metabolic reactions and includes a detailed representation of the ETC and a biomass equation based on experimental measurements. Both computational and experimental approaches were used to investigate light-driven metabolism in Cyanothece sp. ATCC 51142, with a particular focus on reductant production and partitioning within the ETC. The simulation results suggest that growth and metabolic flux distributions are substantially impacted by the relative amounts of light going into the individual photosystems. When growth is limited by the flux through photosystem I, terminal respiratory oxidases are predicted to be an important mechanism for removing excess reductant. Similarly, under photosystem II flux limitation, excess electron carriers must be removed via cyclic electron transport. Furthermore, in silico calculations were in good quantitative agreement with the measured growth rates whereas predictions of reaction usage were qualitatively consistent with protein and mRNA expression data, which we used to further improve the resolution of intracellular flux values.

  13. Effect of bioconversion conditions on vanillin production by Amycolatopsis sp. ATCC 39116 through an analysis of competing by-product formation.

    Science.gov (United States)

    Ma, Xiao-kui; Daugulis, Andrew J

    2014-05-01

    This study investigated the effects of transformation conditions such as initial pH, the initial concentration of glucose and yeast extract in the medium, and the separate addition of ferulic acid and vanillic acid, on the production of vanillin through an analysis of competing by-product formation by Amycolatopsis sp. ATCC 39116. The extent and nature of by-product formation and vanillin yield were affected by initial pH and different initial concentrations of glucose and yeast extract in the medium, with a high yield of vanillin and high cell density obtained at pH 8.0, 10 g/l glucose, and 8 g/l yeast extract. High concentrations of ferulic acid were found to negatively affect cell density. Additional supplementation of 100 mg/l vanillic acid, a metabolically linked by-product, was found to result in a high concentration of vanillin and guaiacol, an intermediate of vanillin. Via an analysis of the effect of these transformation conditions on competing by-product formation, high concentrations of ferulic acid were transformed with a molar yield to vanillin of 96.1 and 95.2 %, by Amycolatopsis sp. ATCC 39116 and Streptomyces V1, respectively, together with a minor accumulation of by-products. These are among the highest performance values reported in the literature to date for Streptomyces in batch cultures. PMID:24078147

  14. Endogenously synthesized (-)-proto-quercitol and glycine betaine are principal compatible solutes of Schizochytrium sp. strain S8 (ATCC 20889) and three new isolates of phylogenetically related thraustochytrids.

    Science.gov (United States)

    Jakobsen, Anita N; Aasen, Inga M; Strøm, Arne R

    2007-09-01

    We report that endogenously synthesized (-)-proto-quercitol (1D-1,3,4/2,5-cyclohexanepentol) and glycine betaine were the principal compatible solutes of Schizochytrium sp. strain S8 (ATCC 20889) and three new osmotolerant isolates of thraustochytrids (strains T65, T66, and T67). The compatible solutes were identified and quantified by use of nuclear magnetic resonance spectroscopy, and their identity was confirmed by mass spectroscopy and measurement of the specific optical rotation. The cellular content of compatible solutes increased with increasing NaCl concentration of a defined medium. (-)-proto-Quercitol was the dominating solute at all NaCl concentrations tested (0.25 to 1.0 M), e.g., cells of S8 and T66 stressed with 1.0 M NaCl accumulated about 500 micromol (-)-proto-quercitol and 100 micromol glycine betaine per g dry weight. To our knowledge, (-)-proto-quercitol has previously been found only in eucalyptus. The 18S rRNA gene sequences of the four (-)-proto-quercitol-producing strains showed 99% identity, and they displayed the same fatty acid profile. The only polyunsaturated fatty acids accumulated were docosahexaenoic acid (78%) and docosapentaenoic acid (22%). A less osmotolerant isolate (strain T29), which was closely phylogenetically related to Thraustochytrium aureum (ATCC 34304), did not contain (-)-proto-quercitol or glycine betaine. Thus, the level of osmotolerance and the osmolyte systems vary among thraustochytrids.

  15. Effect of bioconversion conditions on vanillin production by Amycolatopsis sp. ATCC 39116 through an analysis of competing by-product formation.

    Science.gov (United States)

    Ma, Xiao-kui; Daugulis, Andrew J

    2014-05-01

    This study investigated the effects of transformation conditions such as initial pH, the initial concentration of glucose and yeast extract in the medium, and the separate addition of ferulic acid and vanillic acid, on the production of vanillin through an analysis of competing by-product formation by Amycolatopsis sp. ATCC 39116. The extent and nature of by-product formation and vanillin yield were affected by initial pH and different initial concentrations of glucose and yeast extract in the medium, with a high yield of vanillin and high cell density obtained at pH 8.0, 10 g/l glucose, and 8 g/l yeast extract. High concentrations of ferulic acid were found to negatively affect cell density. Additional supplementation of 100 mg/l vanillic acid, a metabolically linked by-product, was found to result in a high concentration of vanillin and guaiacol, an intermediate of vanillin. Via an analysis of the effect of these transformation conditions on competing by-product formation, high concentrations of ferulic acid were transformed with a molar yield to vanillin of 96.1 and 95.2 %, by Amycolatopsis sp. ATCC 39116 and Streptomyces V1, respectively, together with a minor accumulation of by-products. These are among the highest performance values reported in the literature to date for Streptomyces in batch cultures.

  16. Analysis of the draft genome of Pseudomonas fluorescens ATCC17400 indicates a capacity to take up iron from a wide range of sources, including different exogenous pyoverdines.

    Science.gov (United States)

    Ye, Lumeng; Matthijs, Sandra; Bodilis, Josselin; Hildebrand, Falk; Raes, Jeroen; Cornelis, Pierre

    2014-08-01

    All fluorescent pseudomonads (Pseudomonas aeruginosa, P. putida, P. fluorescens, P. syringae and others) are known to produce the high-affinity peptidic yellow-green fluorescent siderophore pyoverdine. These siderophores have peptide chains that are quite diverse and more than 50 pyoverdine structures have been elucidated. In the majority of the cases, a Pseudomonas species is also able to produce a second siderophore of lower affinity for iron. Pseudomonas fluorescens ATCC 17400 has been shown to produce a unique second siderophore, (thio)quinolobactin, which has an antimicrobial activity against the phytopathogenic Oomycete Pythium debaryanum. We show that this strain has the capacity to utilize 16 different pyoverdines, suggesting the presence of several ferripyoverdine receptors. Analysis of the draft genome of P. fluorescens ATCC 17400 confirmed the presence of 55 TonB-dependent receptors, the largest so far for Pseudomonas, among which 15 are predicted to be ferripyoverdine receptors (Fpv). Phylogenetic analysis revealed the presence of two different clades containing ferripyoverdine receptors, with sequences similar to the P. aeruginosa type II FpvA forming a separate cluster. Among the other receptors we confirmed the presence of the QbsI (thio)quinolobactin receptor, an ferri-achromobactin and an ornicorrugatin receptor, several catecholate and four putative heme receptors. Twenty five of the receptors genes were found to be associated with genes encoding extracytoplasmic sigma factors (ECF σ) and transmembrane anti-σ sensors.

  17. Accurate Localization of the Integration Sites of Two Genomic Islands at Single-Nucleotide Resolution in the Genome of Bacillus cereus ATCC 10987

    Directory of Open Access Journals (Sweden)

    Ren Zhang

    2008-01-01

    Full Text Available We have identified two genomic islands, that is, BCEGI-1 and BCEGI-2, in the genome of Bacillus cereus ATCC 10987, based on comparative analysis with Bacillus cereus ATCC 14579. Furthermore, by using the cumulative GC profile and performing homology searches between the two genomes, the integration sites of the two genomic islands were determined at single-nucleotide resolution. BCEGI-1 is integrated between 159705 bp and 198000 bp, whereas BCEGI-2 is integrated between the end of ORF BCE4594 and the start of the intergenic sequence immediately following BCE4626, that is, from 4256803 bp to 4285534 bp. BCEGI-1 harbors two bacterial Tn7 transposons, which have two sets of genes encoding TnsA, B, C, and D. It is generally believed that unlike the TnsABC+E pathway, the TnsABC+D pathway would only promote vertical transmission to daughter cells. The evidence presented in this paper, however, suggests a role of the TnsABC+D pathway in the horizontal transfer of some genomic islands.

  18. The presence of embedded bacterial pure cultures in agar plates stimulate the culturability of soil bacteria

    DEFF Research Database (Denmark)

    Burmølle, Mette; Johnsen, Kaare; Abu Al-Soud, Waleed Mohamad Abdel F;

    2009-01-01

    cultures in between two layers of agar. Plates containing either embedded Pseudomonas putida or Arthrobacter globiformis resulted in higher numbers of CFUs of soil bacteria (21% and 38%, respectively) after 833 h of incubation, compared to plates with no embedded strain. This indicates a stimulatory effect...... homology to known sequenced isolates in GenBank were recovered from plates with embedded strains than from those without, which indicate a higher number of potential novel soil isolates. This approach for cultivation is therefore a feasible alternative or supplement to traditional cultivation on agar...

  19. Transformation of Synechococcus with a gene for choline oxidase enhances tolerance to salt stress.

    Science.gov (United States)

    Deshnium, P; Los, D A; Hayashi, H; Mustardy, L; Murata, N

    1995-12-01

    Choline oxidase, isolated from the soil bacterium Arthrobacter globiformis, converts choline to glycinebetaine (N-trimethylglycine) without a requirement for any cofactors. The gene for this enzyme, designated codA, was cloned and introduced into the cyanobacterium Synechococcus sp. PCC 7942. The codA gene was expressed under the control of a strong constitutive promoter, and the transformed cells accumulated glycinebetaine at intracellular levels of 60-80 mM. Consequently the cells acquired tolerance to salt stress, as evaluated in terms of growth, accumulation of chlorophyll and photosynthetic activity. PMID:8555454

  20. Comparative proteomic analysis of extracellular secreted proteins expressed by two pathogenic Acanthamoeba castellanii clinical isolates and a non-pathogenic ATCC strain.

    Science.gov (United States)

    Huang, Jian-Ming; Lin, Wei-Chen; Li, Sung-Chou; Shih, Min-Hsiu; Chan, Wen-Ching; Shin, Jyh-Wei; Huang, Fu-Chin

    2016-07-01

    Acanthamoeba keratitis (AK) is a serious ocular disease caused by pathogenic Acanthamoeba gaining entry through wounds in the corneal injury; generally, patients at risk for contracting AK wear contact lenses, usually over a long period of time. Moreover, pathogenic Acanthamoeba causes serious consequences: it makes the cornea turbid and difficult to operate on, including procedures such as enucleation of the eyeball. At present, diagnosis of this disease is not straightforward, and treatment is very demanding. We have established the comparative transcriptome and extracellular secreted proteomic database according to the non-pathogenic strain ATCC 30010 and the pathogenic strains NCKU_B and NCKU_D. We identified 44 secreted proteins successfully, 10 consensus secreted proteins and 34 strain-specific secreted proteins. These proteins may provide targets for therapy and immuno-diagnosis of Acanthamoeba infections. This study shows a suitable approach to identify secreted proteins in Acanthamoeba and provides new perspectives for the study of molecules potentially involved in the AK.

  1. pUCL287 plasmid from Tetragenococcus halophila (Pediococcus halophilus) ATCC 33315 represents a new theta-type replicon family of lactic acid bacteria.

    Science.gov (United States)

    Benachour, A; Frère, J; Novel, G

    1995-05-01

    A cryptic plasmid, pUCL287, was isolated from Tetragenococcus halophila (Pediococcus halophilus) ATCC 33315. It had a theta-type mechanism of replication in its natural host. Its minimal replicon, Rep287, was isolated on a 1.6-kb EcoRI fragment. The Rep287 host range included the genera Pediococcus, Enterococcus, Lactobacillus and Leuconostoc but not genus Lactococcus. Plasmids hybridizing to pUCL287 are rare among lactic acid bacteria. As assessed by hybridization, Rep287 is dissimilar to pAM beta 1, pIP501 and pUCL22, representatives of the most common theta-type replicon groups in Gram-positive bacteria. Therefore, pUCL287 appears to represent a new theta-type replicon family from lactic acid bacteria. PMID:7750734

  2. Increases of ferrous iron oxidation activity and arsenic stressed cell growth by overexpression of Cyc2 in Acidithiobacillus ferrooxidans ATCC19859.

    Science.gov (United States)

    Liu, Wei; Lin, Jianqun; Pang, Xin; Mi, Shuang; Cui, Shuang; Lin, Jianqiang

    2013-01-01

    Acidithiobacillus ferrooxidans plays an important role in bioleaching in reproducing the mineral oxidant of ferric iron (Fe(3+) ) by oxidization of ferrous iron (Fe(2+) ). The high-molecular-weight c-type cytochrome Cyc2 that is located in the external membrane is postulated as the first electron carrier in the Fe(2+) oxidation respiratory pathway of A. ferrooxidans. To increase ferrous iron oxidation activity, a recombinant plasmid pTCYC2 containing cyc2 gene under the control of Ptac promoter was constructed and transferred into A. ferrooxidans ATCC19859. The transcriptional level of cyc2 gene was increased by 2.63-fold and Cyc2 protein expression was observed in the recombinant strain compared with the control. The ferrous iron oxidation activity and the arsenic stressed cell growth of the recombinant strain were also elevated.

  3. Mario Aguilera Peña, coordinador. El orden desarmado. La resistencia de la Asociación de Trabajadores Campesinos del Carare (ATCC

    Directory of Open Access Journals (Sweden)

    Leidy Carolina Navarro Antolínez

    2012-09-01

    Full Text Available Esta investigación representa el doceavo informe del Grupo de Memoria Histórica de la Comisión Nacional de Reparación y Reconciliación (CNRR, que hasta la fecha ha publicado diecinueve investigaciones. El proyecto surgió con la Ley de Justicia y Paz, del año 2005, con el propósito de "presentar informes sobre el origen y evolución de los grupos armados ilegales" (artículo 51. En este informe, el historiador y abogado Mario Aguilera Pena presenta una investigación que, como su titulo lo advierte, tiene por objeto reconstruir la historia de un orden social desarmado, como programa y proceso liderado por la Asociación de Trabajadores y Campesinos del Carare (ATCC, creada en 1987, en el corregimiento de La India, Santander.

  4. Molecular basis for redox-Bohr and cooperative effects in cytochrome c3 from Desulfovibrio desulfuricans ATCC 27774: crystallographic and modelling studies of oxidised and reduced high-resolution structures at pH 7.6

    NARCIS (Netherlands)

    Bento, I.; Matias, P.M.; Baptista, A.M.; Costa, da P.N.; Dongen, van W.M.A.M.; Saraiva, L.M.; Schneider, T.R.; Soares, C.M.; Carrondo, M.A.

    2004-01-01

    The tetraheme cytochrome c, is a small metalloprotein with ca. 13,000 Da found in sulfate-reducing bacteria, which is believed to act as a partner of hydrogenase. The three-dimensional structure of the oxidized and reduced forms of cytochrome c(3) from Desulfovibrio desulfuricans ATCC 27774 at pH 7.

  5. Identification of an L-methionine γ-lyase involved in the production of hydrogen sulfide from L-cysteine in Fusobacterium nucleatum subsp. nucleatum ATCC 25586.

    Science.gov (United States)

    Suwabe, Kyosuke; Yoshida, Yasuo; Nagano, Keiji; Yoshimura, Fuminobu

    2011-10-01

    Fusobacterium nucleatum produces an abundance of hydrogen sulfide (H(2)S) in the oral cavity that is mediated by several enzymes. The identification and characterization of three distinct enzymes (Fn0625, Fn1055 and Fn1220) in F. nucleatum that catalyse the production of H(2)S from l-cysteine have been reported. In the current study, a novel enzyme involved in the production of H(2)S in F. nucleatum ATCC 25586, whose molecular mass had been estimated to be approximately 130 kDa, was identified by two-dimensional electrophoresis combined with MALDI-TOF MS. The enzyme, Fn1419, has previously been characterized as an l-methionine γ-lyase. SDS-PAGE and gel-filtration chromatography indicated that Fn1419 has a molecular mass of 43 kDa and forms tetramers in solution. Unlike other enzymes associated with H(2)S production in F. nucleatum, the quaternary structure of Fn1419 was not completely disrupted by exposure to SDS. The purified recombinant enzyme exhibited a K(m) of 0.32±0.02 mM and a k(cat) of 0.69±0.01 s(-1). Based on current and published data, the enzymic activity for H(2)S production from l-cysteine in F. nucleatum is ranked as follows: Fn1220>Fn1055>Fn1419>Fn0625. Based on kinetic values and relative mRNA levels of the respective genes, as determined by real-time quantitative PCR, the amount of H(2)S produced by Fn1419 was estimated to be 1.9 % of the total H(2)S produced from l-cysteine in F. nucleatum ATCC 25586. In comparison, Fn1220 appeared to contribute significantly to H(2)S production (87.6 %).

  6. Transcriptome sequence and plasmid copy number analysis of the brewery isolate Pediococcus claussenii ATCC BAA-344 T during growth in beer.

    Science.gov (United States)

    Pittet, Vanessa; Phister, Trevor G; Ziola, Barry

    2013-01-01

    Growth of specific lactic acid bacteria in beer leads to spoiled product and economic loss for the brewing industry. Microbial growth is typically inhibited by the combined stresses found in beer (e.g., ethanol, hops, low pH, minimal nutrients); however, certain bacteria have adapted to grow in this harsh environment. Considering little is known about the mechanisms used by bacteria to grow in and spoil beer, transcriptome sequencing was performed on a variant of the beer-spoilage organism Pediococcus claussenii ATCC BAA-344(T) (Pc344-358). Illumina sequencing was used to compare the transcript levels in Pc344-358 growing mid-exponentially in beer to those in nutrient-rich MRS broth. Various operons demonstrated high gene expression in beer, several of which are involved in nutrient acquisition and overcoming the inhibitory effects of hop compounds. As well, genes functioning in cell membrane modification and biosynthesis demonstrated significantly higher transcript levels in Pc344-358 growing in beer. Three plasmids had the majority of their genes showing increased transcript levels in beer, whereas the two cryptic plasmids showed slightly decreased gene expression. Follow-up analysis of plasmid copy number in both growth environments revealed similar trends, where more copies of the three non-cryptic plasmids were found in Pc344-358 growing in beer. Transcriptome sequencing also enabled the addition of several genes to the P. claussenii ATCC BAA-344(T) genome annotation, some of which are putatively transcribed as non-coding RNAs. The sequencing results not only provide the first transcriptome description of a beer-spoilage organism while growing in beer, but they also highlight several targets for future exploration, including genes that may have a role in the general stress response of lactic acid bacteria.

  7. Transcriptome sequence and plasmid copy number analysis of the brewery isolate Pediococcus claussenii ATCC BAA-344 T during growth in beer.

    Directory of Open Access Journals (Sweden)

    Vanessa Pittet

    Full Text Available Growth of specific lactic acid bacteria in beer leads to spoiled product and economic loss for the brewing industry. Microbial growth is typically inhibited by the combined stresses found in beer (e.g., ethanol, hops, low pH, minimal nutrients; however, certain bacteria have adapted to grow in this harsh environment. Considering little is known about the mechanisms used by bacteria to grow in and spoil beer, transcriptome sequencing was performed on a variant of the beer-spoilage organism Pediococcus claussenii ATCC BAA-344(T (Pc344-358. Illumina sequencing was used to compare the transcript levels in Pc344-358 growing mid-exponentially in beer to those in nutrient-rich MRS broth. Various operons demonstrated high gene expression in beer, several of which are involved in nutrient acquisition and overcoming the inhibitory effects of hop compounds. As well, genes functioning in cell membrane modification and biosynthesis demonstrated significantly higher transcript levels in Pc344-358 growing in beer. Three plasmids had the majority of their genes showing increased transcript levels in beer, whereas the two cryptic plasmids showed slightly decreased gene expression. Follow-up analysis of plasmid copy number in both growth environments revealed similar trends, where more copies of the three non-cryptic plasmids were found in Pc344-358 growing in beer. Transcriptome sequencing also enabled the addition of several genes to the P. claussenii ATCC BAA-344(T genome annotation, some of which are putatively transcribed as non-coding RNAs. The sequencing results not only provide the first transcriptome description of a beer-spoilage organism while growing in beer, but they also highlight several targets for future exploration, including genes that may have a role in the general stress response of lactic acid bacteria.

  8. No evidence of harms of probiotic Lactobacillus rhamnosus GG ATCC 53103 in healthy elderly-a phase I open label study to assess safety, tolerability and cytokine responses.

    Directory of Open Access Journals (Sweden)

    Patricia L Hibberd

    Full Text Available BACKGROUND: Although Lactobacillus rhamnosus GG ATCC 53103 (LGG has been consumed by 2 to 5 million people daily since the mid 1990s, there are few clinical trials describing potential harms of LGG, particularly in the elderly. OBJECTIVES: The primary objective of this open label clinical trial is to assess the safety and tolerability of 1×1010 colony forming units (CFU of LGG administered orally twice daily to elderly volunteers for 28 days. The secondary objectives were to evaluate the effects of LGG on the gastrointestinal microbiome, host immune response and plasma cytokines. METHODS: Fifteen elderly volunteers, aged 66-80 years received LGG capsules containing 1×1010 CFU, twice daily for 28 days and were followed through day 56. Volunteers completed a daily diary, a telephone call on study days 3, 7 and 14 and study visits in the Clinical Research Center at baseline, day 28 and day 56 to determine whether adverse events had occurred. Assessments included prompted and open-ended questions. RESULTS: There were no serious adverse events. The 15 volunteers had a total of 47 events (range 1-7 per volunteer, 39 (83% of which were rated as mild and 40% of which were considered related to consuming LGG. Thirty-one (70% of the events were expected, prompted symptoms while 16 were unexpected events. The most common adverse events were gastrointestinal (bloating, gas, and nausea, 27 rated as mild and 3 rated as moderate. In the exploratory analysis, the pro-inflammatory cytokine interleukin 8 decreased during LGG consumption, returning towards baseline one month after discontinuing LGG (p = 0.038 while there was no difference in other pro- or anti-inflammatory plasma cytokines. CONCLUSIONS: Lactobacillus rhamnosus GG ATCC 53103 is safe and well tolerated in healthy adults aged 65 years and older. TRIAL REGISTRATION: ClinicalTrials.gov NCT 01274598.

  9. Proteome Analyses of Strains ATCC 51142 and PCC 7822 of the Diazotrophic Cyanobacterium Cyanothece sp under Culture Conditions Resulting in Enhanced H-2 Production

    Energy Technology Data Exchange (ETDEWEB)

    Aryal, Uma K.; Callister, Stephen J.; Mishra, Sujata; Zhang, Xiaohui; Shutthanandan, Janani I.; Angel, Thomas E.; Shukla, Anil K.; Monroe, Matthew E.; Moore, Ronald J.; Koppenaal, David W.; Smith, Richard D.; Sherman, Louis

    2013-02-01

    Cultures of the cyanobacterial genus Cyanothece have been shown to produce high levels of biohydrogen. These strains are diazotrophic and undergo pronounced diurnal cycles when grown under N2-fixing conditions in light-dark cycles. We seek to better understand the way in which proteins respond to these diurnal changes and we performed quantitative proteome analysis of Cyanothece ATCC 51142 and PCC 7822 grown under 8 different nutritional conditions. Nitrogenase expression was limited to N2-fixing conditions, and in the absence of glycerol, nitrogenase gene expression was linked to the dark period. However, glycerol induced expression of nitrogenase during part of the light period, together with cytochrome c oxidase (Cox), glycogen phosphorylase (Glp), and glycolytic and pentose-phosphate pathway (PPP) enzymes. This indicated that nitrogenase expression in the light was facilitated via higher respiration and glycogen breakdown. Key enzymes of the Calvin cycle were inhibited in Cyanothece ATCC 51142 in the presence of glycerol under H2 producing conditions, suggesting a competition between these sources of carbon. However, in Cyanothece PCC 7822, the Calvin cycle still played a role in cofactor recycling during H2 production. Our data comprise the first comprehensive profiling of proteome changes in Cyanothece PCC 7822, and allows an in-depth comparative analysis of major physiological and biochemical processes that influence H2-production in both the strains. Our results revealed many previously uncharacterized proteins that may play a role in nitrogenase activity and in other metabolic pathways and may provide suitable targets for genetic manipulation that would lead to improvement of large scale H2 production.

  10. 蜡状芽孢杆菌ATCC 10987基因组蛋白质编码基因的重新注释与分析%Re-annotation and analysis of protein-coding genes in Bacillus cereus ATCC 10987 genome

    Institute of Scientific and Technical Information of China (English)

    林岩

    2007-01-01

    蜡状芽胞杆菌ATCC 10987对于细菌比较基因组学的研究有重要意义.但其染色体中基因的数目被RefSeq注释为5 603个,这个注释是有疑问的.本文采用Zcurve和Glimmer程序联合打分的方法来识别其蛋白质编码基因.为保证预测结果的可靠性,对联合判别附加预测的基因使用了BLAST方法进行数据库同源性搜索.结果,蜡状芽胞杆菌ATCC 10987基因组中的蛋白质编码基因的数目被重新确定为5 180个.这个数目明显低于原始注释的数目,并且一些指标表明新的注释更为可信.这些相对正确的基因集合为该细菌亲缘物种的深入研究提供了基础.

  11. Diversity and transcription of proteases involved in the maturation of hydrogenases in Nostoc punctiforme ATCC 29133 and Nostoc sp. strain PCC 7120

    Directory of Open Access Journals (Sweden)

    Lindblad Peter

    2009-03-01

    Full Text Available Abstract Background The last step in the maturation process of the large subunit of [NiFe]-hydrogenases is a proteolytic cleavage of the C-terminal by a hydrogenase specific protease. Contrary to other accessory proteins these hydrogenase proteases are believed to be specific whereby one type of hydrogenases specific protease only cleaves one type of hydrogenase. In cyanobacteria this is achieved by the gene product of either hupW or hoxW, specific for the uptake or the bidirectional hydrogenase respectively. The filamentous cyanobacteria Nostoc punctiforme ATCC 29133 and Nostoc sp strain PCC 7120 may contain a single uptake hydrogenase or both an uptake and a bidirectional hydrogenase respectively. Results In order to examine these proteases in cyanobacteria, transcriptional analyses were performed of hupW in Nostoc punctiforme ATCC 29133 and hupW and hoxW in Nostoc sp. strain PCC 7120. These studies revealed numerous transcriptional start points together with putative binding sites for NtcA (hupW and LexA (hoxW. In order to investigate the diversity and specificity among hydrogeanse specific proteases we constructed a phylogenetic tree which revealed several subgroups that showed a striking resemblance to the subgroups previously described for [NiFe]-hydrogenases. Additionally the proteases specificity was also addressed by amino acid sequence analysis and protein-protein docking experiments with 3D-models derived from bioinformatic studies. These studies revealed a so called "HOXBOX"; an amino acid sequence specific for protease of Hox-type which might be involved in docking with the large subunit of the hydrogenase. Conclusion Our findings suggest that the hydrogenase specific proteases are under similar regulatory control as the hydrogenases they cleave. The result from the phylogenetic study also indicates that the hydrogenase and the protease have co-evolved since ancient time and suggests that at least one major horizontal gene transfer

  12. 发酵产菊糖果糖转移酶的培养基设计及菌体生长动力学研究%Optimization of Inulin Fructotransferase-producing Medium and Determination of Cell Growth Kinetic Parameter for Arthrobacter ureafaciens SK-8.001

    Institute of Scientific and Technical Information of China (English)

    卢峥嵘; 赵萌; 沐万孟; 张涛; 江波

    2009-01-01

    In this paper,the optimal inulin fructotransferase-producing medium and fermentation kinetic parameters of Arthrobacter ureafaciens SK -8.001 were studied. The recipes of optimal media included 2. 5% inulin,0. 3% NaN0_3,0. 1% yeast extract,0. 01% MgSO_4,0. 04% KH_2P0_4,0. 001% FeSO_4·7H_2O. The maximum enzyme activity reached 19. 85 U/mL,which was 2. 04 fold higher than in initial media. The study on the fermentation process in the optimal media suggested the right time to add inducers into the media was 12 h or 66th hours during the fermentation. The cell growth kinetic model for SK-8.001 in batch fermentation was X (t) = 9. 356 05/ [1 + exp~((4.4222-0.1102l)) ].%对1株产菊糖果糖转移酶(inulin fructotransferase)的金黄色节杆菌(Arthrobacter ureafaciens)SK-8.001的产酶培养基进行了优化,同时对其在分批发酵过程中培养基组分变化和菌体生长动力学进行了研究.所得到的改进型产酶培养基配方为25 g/L菊糖,3 g/L NaNO_3,1g/L酵母膏,0.1 g/L MgSO_4,0.4g/L KH_2PO_4,0.01g/LFeSO_4·7H_2O.在此培养基中的最高酶活达到19.85 U/mL,较初始培养基提高了2.04倍.对在此培养基中Sk-8.001的发酵过程研究发现发酵12 h和66 h是较为理想的诱导物添加时刻.得到SK-8.001在优化培养基中分批培养的菌体生长动力学模型为:X(t)=9.356 05/[1+exp~((4.4222-0.1102:))].

  13. The Effects of Heterologous Expression of groEL from Arthrobacter simplex on the Resistance Performance of Escherichia coli%简单节杆菌groEL基因表达对大肠杆菌抗逆性能的影响

    Institute of Scientific and Technical Information of China (English)

    王艳霞; 王红玲; 王敏; 骆健美

    2016-01-01

    Arthrobacter simplex,a strain with steroid C1,2 dehydrogenation ability,has been widely used in the steroid industry, usually adding ethanol in the transforming system may catalyze the dissolution of hydrophobic substrates. Our prior work by LC-MS/MS and bioinformatics analysis found that heat shock protein(HSP)GroEL was closely correlated to the cell adaptive behavior under ethanol stress condition. Based on this concept,groEL gene was cloned by PCR while usingA. simplexTCCC 11037 genome as a template. The results showed that the homologies of nucleotide and amino acid sequence of GroEL in the strain were the highest with that fromNocardioides sp. JS614 by the ratio of 89% and 95%,respectively. Then its heterologous expression was conducted in Escherichia coli BL21(DE3)with the vector pET-22b (+),and the results showed that the recombinant strain presented the significant increase on the survival performance under the shock condition with high-concentration of methanol,hypertonicity and superacid,as well as the growth performance under the proper stresses of them. The study was the first report on the cloning and expression of genegroEL fromA. simplex,and also provided fundamental data for exploring the biological function of HSP protein GroEL from A. simplex.%以简单节杆菌(Arthrobacter simplex)TCCC 11037基因组为模板,通过PCR方法进行groEL基因克隆,结果表明,该菌株中GroEL的核苷酸和氨基酸序列与Nocardioidessp. JS614来源的GroEL同源性最高,分别为89%和95%。通过pET-22b(+)将其在大肠杆菌BL21(DE3)中进行异源表达,结果表明,重组菌株在甲醇、高渗和强酸这3种压力高浓度冲击条件下的存活能力和适当压力条件下的生长性能均明显提高。

  14. Cloning and sequencing of the kedarcidin biosynthetic gene cluster from Streptoalloteichus sp. ATCC 53650 revealing new insights into biosynthesis of the enediyne family of antitumor antibiotics†

    Science.gov (United States)

    Lohman, Jeremy R.; Huang, Sheng-Xiong; Horsman, Geoffrey P.; Dilfer, Paul E.; Huang, Tingting; Chen, Yihua; Wendt-Pienkowski, Evelyn; Shen, Ben

    2013-01-01

    Enediyne natural product biosynthesis is characterized by a convergence of multiple pathways, generating unique peripheral moieties that are appended onto the distinctive enediyne core. Kedarcidin (KED) possesses two unique peripheral moieties, a (R)-2-aza-3-chloro-β-tyrosine and an iso-propoxy-bearing 2-naphthonate moiety, as well as two deoxysugars. The appendage pattern of these peripheral moieties to the enediyne core in KED differs from the other enediynes studied to date with respect to stereochemical configuration. To investigate the biosynthesis of these moieties and expand our understanding of enediyne core formation, the biosynthetic gene cluster for KED was cloned from Streptoalloteichus sp. ATCC 53650 and sequenced. Bioinformatics analysis of the ked cluster revealed the presence of the conserved genes encoding for enediyne core biosynthesis, type I and type II polyketide synthase loci likely responsible for 2-aza-L-tyrosine and 3,6,8-trihydroxy-2-naphthonate formation, and enzymes known for deoxysugar biosynthesis. Genes homologous to those responsible for the biosynthesis, activation, and coupling of the L-tyrosine-derived moieties from C-1027 and maduropeptin and of the naphthonate moiety from neocarzinostatin are present in the ked cluster, supporting 2-aza-L-tyrosine and 3,6,8-trihydroxy-2-naphthoic acid as precursors, respectively, for the (R)-2-aza-3-chloro-β-tyrosine and the 2-naphthonate moieties in KED biosynthesis. PMID:23360970

  15. Cloning and sequencing of the kedarcidin biosynthetic gene cluster from Streptoalloteichus sp. ATCC 53650 revealing new insights into biosynthesis of the enediyne family of antitumor antibiotics.

    Science.gov (United States)

    Lohman, Jeremy R; Huang, Sheng-Xiong; Horsman, Geoffrey P; Dilfer, Paul E; Huang, Tingting; Chen, Yihua; Wendt-Pienkowski, Evelyn; Shen, Ben

    2013-03-01

    Enediyne natural product biosynthesis is characterized by a convergence of multiple pathways, generating unique peripheral moieties that are appended onto the distinctive enediyne core. Kedarcidin (KED) possesses two unique peripheral moieties, a (R)-2-aza-3-chloro-β-tyrosine and an iso-propoxy-bearing 2-naphthonate moiety, as well as two deoxysugars. The appendage pattern of these peripheral moieties to the enediyne core in KED differs from the other enediynes studied to date with respect to stereochemical configuration. To investigate the biosynthesis of these moieties and expand our understanding of enediyne core formation, the biosynthetic gene cluster for KED was cloned from Streptoalloteichus sp. ATCC 53650 and sequenced. Bioinformatics analysis of the ked cluster revealed the presence of the conserved genes encoding for enediyne core biosynthesis, type I and type II polyketide synthase loci likely responsible for 2-aza-l-tyrosine and 3,6,8-trihydroxy-2-naphthonate formation, and enzymes known for deoxysugar biosynthesis. Genes homologous to those responsible for the biosynthesis, activation, and coupling of the l-tyrosine-derived moieties from C-1027 and maduropeptin and of the naphthonate moiety from neocarzinostatin are present in the ked cluster, supporting 2-aza-l-tyrosine and 3,6,8-trihydroxy-2-naphthoic acid as precursors, respectively, for the (R)-2-aza-3-chloro-β-tyrosine and the 2-naphthonate moieties in KED biosynthesis. PMID:23360970

  16. Sequence Analysis of Inducible Prophage phIS3501 Integrated into the Haemolysin II Gene of Bacillus thuringiensis var israelensis ATCC35646

    Directory of Open Access Journals (Sweden)

    Bouziane Moumen

    2012-01-01

    Full Text Available Diarrheic food poisoning by bacteria of the Bacillus cereus group is mostly due to several toxins encoded in the genomes. One of them, cytotoxin K, was recently identified as responsible for severe necrotic syndromes. Cytotoxin K is similar to a class of proteins encoded by genes usually annotated as haemolysin II (hlyII in the majority of genomes of the B. cereus group. The partially sequenced genome of Bacillus thuringiensis var israelensis ATCC35646 contains several potentially induced prophages, one of them integrated into the hlyII gene. We determined the complete sequence and established the genomic organization of this prophage-designated phIS3501. During induction of excision of this prophage with mitomycin C, intact hlyII gene is formed, thus providing to cells a genetic ability to synthesize the active toxin. Therefore, this prophage, upon its excision, can be implicated in the regulation of synthesis of the active toxin and thus in the virulence of bacterial host. A generality of selection for such systems in bacterial pathogens is indicated by the similarity of this genetic arrangement to that of Staphylococcus aureus  β-haemolysin.

  17. Bacillus cereus ATCC 14579 RpoN (Sigma 54) Is a Pleiotropic Regulator of Growth, Carbohydrate Metabolism, Motility, Biofilm Formation and Toxin Production.

    Science.gov (United States)

    Hayrapetyan, Hasmik; Tempelaars, Marcel; Nierop Groot, Masja; Abee, Tjakko

    2015-01-01

    Sigma 54 is a transcriptional regulator predicted to play a role in physical interaction of bacteria with their environment, including virulence and biofilm formation. In order to study the role of Sigma 54 in Bacillus cereus, a comparative transcriptome and phenotypic study was performed using B. cereus ATCC 14579 WT, a markerless rpoN deletion mutant, and its complemented strain. The mutant was impaired in many different cellular functions including low temperature and anaerobic growth, carbohydrate metabolism, sporulation and toxin production. Additionally, the mutant showed lack of motility and biofilm formation at air-liquid interphase, and this correlated with absence of flagella, as flagella staining showed only WT and complemented strain to be highly flagellated. Comparative transcriptome analysis of cells harvested at selected time points during growth in aerated and static conditions in BHI revealed large differences in gene expression associated with loss of phenotypes, including significant down regulation of genes in the mutant encoding enzymes involved in degradation of branched chain amino acids, carbohydrate transport and metabolism, flagella synthesis and virulence factors. Our study provides evidence for a pleiotropic role of Sigma 54 in B. cereus supporting its adaptive response and survival in a range of conditions and environments.

  18. Tamarind seed powder and palm kernel cake: two novel agro residues for the production of tannase under solid state fermentation by Aspergillus niger ATCC 16620.

    Science.gov (United States)

    Sabu, A; Pandey, A; Daud, M Jaafar; Szakacs, G

    2005-07-01

    Palm kernel cake (PKC), the residue obtained after extraction of palm oil from oil palm seeds and tamarind seed powder (TSP) obtained after removing the fruit pulp from tamarind fruit pod were tested for the production of tannase under solid-state fermentation (SSF) using Aspergillus niger ATCC 16620. The fungal strain was grown on the substrates without any pretreatment. In PKC medium, a maximum enzyme yield of 13.03 IU/g dry substrate (gds) was obtained when SSF was carried out at 30 degrees C, 53.5% initial substrate moisture, 33 x 10(9) spores/5 g substrate inoculum size and 5% tannic acid as additional carbon source after 96 h of fermentation. In TSP medium, maximum tannase yield of 6.44 IU/gds was obtained at 30 degrees C, 65.75% initial substrate moisture, 11 x 10(9) spores/5 g substrate inoculum, 1% glycerol as additional carbon source and 1% potassium nitrate as additional nitrogen source after 120 h of fermentation. Results from the study are promising for the economic utilization and value addition of these important agro residues, which are abundantly available in many tropical and subtropical countries.

  19. Myo-inositol hexakisphosphate degradation by Bifidobacterium pseudocatenulatum ATCC 27919 improves mineral availability of high fibre rye-wheat sour bread.

    Science.gov (United States)

    García-Mantrana, Izaskun; Monedero, Vicente; Haros, Monika

    2015-07-01

    The goal of this investigation was to develop baking products using Bifidobacterium pseudocatenulatum ATCC27919, a phytase producer, as a starter in sourdough for the production of whole rye-wheat mixed bread. This Bifidobacterium strain contributed to myo-inositol hexakisphosphate (phytate) hydrolysis, resulting in breads with higher mineral availability as was predicted by the phytate/mineral molar ratios, which remained below the inhibitory threshold values for Ca and Zn intestinal absorption. The products with sourdough showed similar technological quality as their homologous without sourdough, with levels of acetic and d/l lactic acids in dough and bread baking significantly higher with the use of sourdough. The overall acceptability scores showed that breads with 25% of whole rye flour were highly accepted regardless of the inclusion of sourdough. This work emphasises that the in situ production of phytase during fermentation by GRAS/QPS microorganisms constitutes a strategy which is particularly appropriate for reducing the phytate contents in products for human consumption.

  20. Determination of the folate content in cladodes of nopal (Opuntia ficus indica) by microbiological assay utilizing Lactobacillus casei (ATCC 7469) and enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Ortiz-Escobar, Tania Breshkovskaya; Valverde-González, Maria Elena; Paredes-López, Octavio

    2010-05-26

    Prickly pear cactus has been an important food source in Mexico since ancient times due to its economical and ecological benefits and potential nutraceutical value. Nevertheless, studies on the nutritional aspects and health benefits have been scarce. The purpose of this study was to assess, apparently for the first time, the folate contents of cladodes of nopal by a microbiological assay, using Lactobacillus casei (ATCC 7469) in extracts that were enzymatically treated to release the bound vitamin, employing single, dual, and trienzymatic procedures, and using the enzyme-linked immunosorbent assay (ELISA). We used Opuntia cladodes of different length sizes. The microbiological assay showed some differences among enzyme treatments and sizes of nopal; the trienzyme treatment (alpha-amylase-protease-conjugase) was more efficient in determining the folate content in nopal, giving 5.0 ng/g in the small size cladodes at 54 h of testing time, while ELISA showed no significant differences in the folate content among sizes of cladodes (5.5-5.62 ng/g at 0 min testing time). Both techniques may be used for the assessment of folate content in cladodes, but ELISA is more rapid and reliable. PMID:20441169

  1. Genome mining of the sordarin biosynthetic gene cluster from Sordaria araneosa Cain ATCC 36386: characterization of cycloaraneosene synthase and GDP-6-deoxyaltrose transferase.

    Science.gov (United States)

    Kudo, Fumitaka; Matsuura, Yasunori; Hayashi, Takaaki; Fukushima, Masayuki; Eguchi, Tadashi

    2016-07-01

    Sordarin is a glycoside antibiotic with a unique tetracyclic diterpene aglycone structure called sordaricin. To understand its intriguing biosynthetic pathway that may include a Diels-Alder-type [4+2]cycloaddition, genome mining of the gene cluster from the draft genome sequence of the producer strain, Sordaria araneosa Cain ATCC 36386, was carried out. A contiguous 67 kb gene cluster consisting of 20 open reading frames encoding a putative diterpene cyclase, a glycosyltransferase, a type I polyketide synthase, and six cytochrome P450 monooxygenases were identified. In vitro enzymatic analysis of the putative diterpene cyclase SdnA showed that it catalyzes the transformation of geranylgeranyl diphosphate to cycloaraneosene, a known biosynthetic intermediate of sordarin. Furthermore, a putative glycosyltransferase SdnJ was found to catalyze the glycosylation of sordaricin in the presence of GDP-6-deoxy-d-altrose to give 4'-O-demethylsordarin. These results suggest that the identified sdn gene cluster is responsible for the biosynthesis of sordarin. Based on the isolated potential biosynthetic intermediates and bioinformatics analysis, a plausible biosynthetic pathway for sordarin is proposed. PMID:27072286

  2. Novel α-L-arabinofuranosidase from Cellulomonas fimi ATCC 484 and its substrate-specificity analysis with the aid of computer.

    Science.gov (United States)

    Yang, Ying; Zhang, Lujia; Guo, Mingrong; Sun, Jiaqi; Matsukawa, Shingo; Xie, Jingli; Wei, Dongzhi

    2015-04-15

    In the process of gene mining for novel α-L-arabinofuranosidases (AFs), the gene Celf_3321 from Cellulomonas fimi ATCC 484 encodes an AF, termed as AbfCelf, with potent activity, 19.4 U/mg under the optimum condition, pH 6.0 and 40 °C. AbfCelf can hydrolyze α-1,5-linked oligosaccharides, sugar beet arabinan, linear 1,5-α-arabinan, and wheat flour arabinoxylan, which is partly different from some previously well-characterized GH 51 AFs. The traditional substrate-specificity analysis for AFs is labor-consuming and money costing, because the substrates include over 30 kinds of various 4-nitrophenol (PNP)-glycosides, oligosaccharides, and polysaccharides. Hence, a preliminary structure and mechanism based method was applied for substrate-specificity analysis. The binding energy (ΔG, kcal/mol) obtained by docking suggested the reaction possibility and coincided with the experimental results. AbfA crystal 1QW9 was used to test the rationality of docking method in simulating the interaction between enzyme and substrate, as well the credibility of the substrate-specificity analysis method in silico. PMID:25797391

  3. Internalization of the PDZ and its photodynamic effect on the growth of ATCC and clinical strains of E. coli and S. aureus

    Science.gov (United States)

    Rodrigues da Silva, Gislene; Henrique Correia Pereira, André; Guerra Pinto, Juliana; José Raniero, Leandro; Ferreira-Strixino, Juliana

    2016-09-01

    The treatment of bacterial infections has been a challenge after the end of the ‘era of antibiotics’. Bacteria are capable of causing many infectious diseases; therefore, with the increasing number of bacteria becoming resistant, development of alternative therapies is needed to minimize, or even eliminate the use of antibiotics. Photodynamic therapy (PDT) is a promising alternative to fight microorganism. In view of the increasing emergence of resistant bacteria and the limitations of conventional treatment, this study evaluated the effect of photodynamic therapy with photodithazine (PDZ) in inactivating bacterial strains of E. coli and S. aureus in vitro, comparing the behavior of clinical and ATCC strains. Confocal microscopy analysis was performed to determine the internalization of the PS and spectrophotometric technique was used to determine the growth of bacteria in vitro. PDT using PDZ was able to reduce the growth of S. aureus strains using the incubation time of 24 h, whereas no satisfactory results were obtained with 15 min incubation. The E. coli strains, tested at two incubation times, did not affectively reduce bacterial growth. Therefore, it is concluded that PDT using PDZ is viable when applied to the S. aureus strains, when suitable incubation times are used.

  4. Molecular analysis of the replication region of the theta-replicating plasmid pUCL287 from Tetragenococcus (Pediococcus) halophilus ATCC33315.

    Science.gov (United States)

    Benachour, A; Frère, J; Flahaut, S; Novel, G; Auffray, Y

    1997-08-01

    The complete nucleotide sequence of the 8.7-kb theta-replicating plasmid pUCL287 from Tetragenococcus halophilus (formerly Pediococcus halophilus) ATCC33315 has been determined. The replication region was identified and analyzed. Its nucleotide sequence contains an untranslated region, the replication origin, followed by two open reading frames (ORFs) encoding two proteins of 311 (RepA287) and 168 (RepB287) amino acids, respectively. Evidence is presented to show that RepA287 represents the plasmid replication protein. RepB287, which is non-essential for replication, is involved in the plasmid copy-number control and segregational stability. The roles of lactococcal proteins homologous to RepB287 have not been defined so far. Nevertheless, the structural organization of the pUCL287 replication region is remarkably similar to those of well known theta-replicating lactococcal plasmids despite the absence of homology of the replication origin and of the replication protein, and this suggests that pUCL287 uses the same mechanism of replication. Nucleotide sequence comparisons show that pSMB74, a pediococcal plasmid encoding bacteriocin production, is a member of the pUCL287 replicon family. PMID:9294035

  5. Effect of C/N Ratio and Media Optimization through Response Surface Methodology on Simultaneous Productions of Intra- and Extracellular Inulinase and Invertase from Aspergillus niger ATCC 20611

    Directory of Open Access Journals (Sweden)

    Mojdeh Dinarvand

    2013-01-01

    Full Text Available The study is to identify the extraction of intracellular inulinase (exo- and endoinulinase and invertase as well as optimization medium composition for maximum productions of intra- and extracellular enzymes from Aspergillus niger ATCC 20611. From two different methods for extraction of intracellular enzymes, ultrasonic method was found more effective. Response surface methodology (RSM with a five-variable and three-level central composite design (CCD was employed to optimize the medium composition. The effect of five main reaction parameters including sucrose, yeast extract, NaNO3, Zn+2, and Triton X-100 on the production of enzymes was analyzed. A modified quadratic model was fitted to the data with a coefficient of determination (R2 more than 0.90 for all responses. The intra-extracellular inulinase and invertase productions increased in the range from 16 to 8.4 times in the optimized medium (10% (w/v sucrose, 2.5% (w/v yeast extract, 2% (w/v NaNO3, 1.5 mM (v/v Zn+2, and 1% (v/v Triton X-100 by RSM and from around 1.2 to 1.3 times greater than in the medium optimized by one-factor-at-a-time, respectively. The results of bioprocesses optimization can be useful in the scale-up fermentation and food industry.

  6. Genome mining of the sordarin biosynthetic gene cluster from Sordaria araneosa Cain ATCC 36386: characterization of cycloaraneosene synthase and GDP-6-deoxyaltrose transferase.

    Science.gov (United States)

    Kudo, Fumitaka; Matsuura, Yasunori; Hayashi, Takaaki; Fukushima, Masayuki; Eguchi, Tadashi

    2016-07-01

    Sordarin is a glycoside antibiotic with a unique tetracyclic diterpene aglycone structure called sordaricin. To understand its intriguing biosynthetic pathway that may include a Diels-Alder-type [4+2]cycloaddition, genome mining of the gene cluster from the draft genome sequence of the producer strain, Sordaria araneosa Cain ATCC 36386, was carried out. A contiguous 67 kb gene cluster consisting of 20 open reading frames encoding a putative diterpene cyclase, a glycosyltransferase, a type I polyketide synthase, and six cytochrome P450 monooxygenases were identified. In vitro enzymatic analysis of the putative diterpene cyclase SdnA showed that it catalyzes the transformation of geranylgeranyl diphosphate to cycloaraneosene, a known biosynthetic intermediate of sordarin. Furthermore, a putative glycosyltransferase SdnJ was found to catalyze the glycosylation of sordaricin in the presence of GDP-6-deoxy-d-altrose to give 4'-O-demethylsordarin. These results suggest that the identified sdn gene cluster is responsible for the biosynthesis of sordarin. Based on the isolated potential biosynthetic intermediates and bioinformatics analysis, a plausible biosynthetic pathway for sordarin is proposed.

  7. Sugarcane molasses and yeast powder used in the Fructooligosaccharides production by Aspergillus japonicus-FCL 119T and Aspergillus niger ATCC 20611.

    Science.gov (United States)

    Dorta, Claudia; Cruz, Rubens; de Oliva-Neto, Pedro; Moura, Danilo José Camargo

    2006-12-01

    Different concentrations of sucrose (3-25% w/v) and peptone (2-5% w/v) were studied in the formulation of media during the cultivation of Aspergillus japonicus-FCL 119T and Aspergillus niger ATCC 20611. Moreover, cane molasses (3.5-17.5% w/v total sugar) and yeast powder (1.5-5% w/v) were used as alternative nutrients for both strains' cultivation. These media were formulated for analysis of cellular growth, beta-Fructosyltransferase and Fructooligosaccharides (FOS) production. Transfructosylating activity (U ( t )) and FOS production were analyzed by HPLC. The highest enzyme production by both the strains was 3% (w/v) sucrose and 3% (w/v) peptone, or 3.5% (w/v) total sugars present in cane molasses and 1.5% (w/v) yeast powder. Cane molasses and yeast powder were as good as sucrose and peptone in the enzyme and FOS (around 60% w/w) production by studied strains. PMID:16835781

  8. Determination of the folate content in cladodes of nopal (Opuntia ficus indica) by microbiological assay utilizing Lactobacillus casei (ATCC 7469) and enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Ortiz-Escobar, Tania Breshkovskaya; Valverde-González, Maria Elena; Paredes-López, Octavio

    2010-05-26

    Prickly pear cactus has been an important food source in Mexico since ancient times due to its economical and ecological benefits and potential nutraceutical value. Nevertheless, studies on the nutritional aspects and health benefits have been scarce. The purpose of this study was to assess, apparently for the first time, the folate contents of cladodes of nopal by a microbiological assay, using Lactobacillus casei (ATCC 7469) in extracts that were enzymatically treated to release the bound vitamin, employing single, dual, and trienzymatic procedures, and using the enzyme-linked immunosorbent assay (ELISA). We used Opuntia cladodes of different length sizes. The microbiological assay showed some differences among enzyme treatments and sizes of nopal; the trienzyme treatment (alpha-amylase-protease-conjugase) was more efficient in determining the folate content in nopal, giving 5.0 ng/g in the small size cladodes at 54 h of testing time, while ELISA showed no significant differences in the folate content among sizes of cladodes (5.5-5.62 ng/g at 0 min testing time). Both techniques may be used for the assessment of folate content in cladodes, but ELISA is more rapid and reliable.

  9. Bacillus cereus ATCC 14579 RpoN (Sigma 54 Is a Pleiotropic Regulator of Growth, Carbohydrate Metabolism, Motility, Biofilm Formation and Toxin Production.

    Directory of Open Access Journals (Sweden)

    Hasmik Hayrapetyan

    Full Text Available Sigma 54 is a transcriptional regulator predicted to play a role in physical interaction of bacteria with their environment, including virulence and biofilm formation. In order to study the role of Sigma 54 in Bacillus cereus, a comparative transcriptome and phenotypic study was performed using B. cereus ATCC 14579 WT, a markerless rpoN deletion mutant, and its complemented strain. The mutant was impaired in many different cellular functions including low temperature and anaerobic growth, carbohydrate metabolism, sporulation and toxin production. Additionally, the mutant showed lack of motility and biofilm formation at air-liquid interphase, and this correlated with absence of flagella, as flagella staining showed only WT and complemented strain to be highly flagellated. Comparative transcriptome analysis of cells harvested at selected time points during growth in aerated and static conditions in BHI revealed large differences in gene expression associated with loss of phenotypes, including significant down regulation of genes in the mutant encoding enzymes involved in degradation of branched chain amino acids, carbohydrate transport and metabolism, flagella synthesis and virulence factors. Our study provides evidence for a pleiotropic role of Sigma 54 in B. cereus supporting its adaptive response and survival in a range of conditions and environments.

  10. Efeito e modo de ação das bacteriocinas produzidas por Lactococcus lactis subsp. lactis ITAL 383, ATCC 11454 e CNRZ 150 contra Listeria innocua LIN 11 Effect and mode of action of the bacterioncin produced by Lactococcus. lactis subsp. lactis ITAL 383, ATCC 11454 e CNRZ 150 against Listeria innocua LIN 11

    Directory of Open Access Journals (Sweden)

    Izildinha MORENO

    1999-01-01

    Full Text Available O efeito e o modo de ação das bacteriocinas produzidas por L. lactis subsp. lactis ITAL 383 e CNRZ 150 são similares à nisina de L. lactis subsp. lactis ATCC 11454. Estas bacteriocinas apresentaram um modo de ação bactericida, causando a lise de células de L. innocua LIN 11, associada ao decréscimo da absorbância e da viabilidade celular. O efeito letal foi maior para células em fase exponencial comparativamente à fase estacionária de crescimento. A adsorção dessas bacteriocinas às células de L. innocua LIN 11 foi muito rápida e influenciada pelo pH do meio de suspensão; adsorção máxima foi verificada a pH 6,0 e logo após o contato inicial. Perda completa de adsorção ocorreu em pH 2,0.The effect and mode of action of the bacteriocin produced by L. lactis subsp. lactis ITAL 383 and CNRZ 150 are similar to the nisin produced by L. lactis subsp. lactis ATCC 11454. It was clearly bactericidal, and caused lysis of a strain of L. innocua LIN 11 detected by the decrease of absorbance values and the cell viability. Their lethal effect was considerably higher during the logarithmic growth when compared to the stationary phase. Adsorption developed rapidly and was influenced by the pH value of the suspension medium. Maximum adsorption was observed at pH 6,0 and immediately after initial contact and loss at pH 2,0.

  11. Effects of bilimbi (Averrhoa bilimbi L.) and tamarind (Tamarindus indica L.) juice on Listeria monocytogenes Scott A and Salmonella Typhimurium ATCC 14028 and the sensory properties of raw shrimps.

    Science.gov (United States)

    Norhana, M N Wan; Azman, Mohd Nor A; Poole, Susan E; Deeth, Hilton C; Dykes, Gary A

    2009-11-30

    The potential of using juice of bilimbi (Averrhoa bilimbi L.) and tamarind (Tamarindus indica L.) to reduce Listeria monocytogenes Scott A and Salmonella Typhimurium ATCC 14028 populations on raw shrimps after washing and during storage (4 degrees C) was investigated. The uninoculated raw shrimps and those inoculated with approximately 9 log cfu/ml of L. monocytogenes Scott A and S. Typhimurium ATCC 14028 were washed (dipped or rubbed) in distilled water (SDW) (control), bilimbi or tamarind juice at 1:4 (w/v) concentrations for 10 and 5 min. Naturally occurring aerobic bacteria (APC), L. monocytogenes Scott A and S. Typhimurium ATCC 14028 counts, pH values and sensory analysis of washed shrimps were determined immediately after washing (day 0), and on days 3 and 7 of storage. Compared to SDW, bilimbi and tamarind juice significantly (p0.05) throughout the storage except for odour (p<0.05) attributes at 0 day when acidic or lemony smell was noticed in bilimbi- and tamarind-washed shrimps and not in control shrimps.

  12. RT-qPCR analysis of putative beer-spoilage gene expression during growth of Lactobacillus brevis BSO 464 and Pediococcus claussenii ATCC BAA-344(T) in beer.

    Science.gov (United States)

    Bergsveinson, Jordyn; Pittet, Vanessa; Ziola, Barry

    2012-10-01

    Lactic acid bacteria (LAB) contamination of beer presents a continual economic threat to brewers. Interestingly, only certain isolates of LAB can grow in the hostile beer environment (e.g., as studied here, Lactobacillus brevis BSO 464 (Lb464) and a non-ropy isolate of Pediococcus claussenii ATCC BAA-344(T) (Pc344NR)), indicating that significant genetic specialization is required. The genes hitA, horA, horB, horC, and bsrA, which have been proposed to confer beer-spoiling ability to an organism, are suspected of counteracting the antimicrobial effects of hops. However, these genes are not present in the same combination (if at all) across beer-spoiling organisms. As such, we sought to investigate the extent to which these genes participate during Lb464 and Pc344NR mid-logarithmic growth in beer through reverse transcription quantitative PCR analysis. We first determined the optimal reference gene set needed for data normalization and, for each bacterium, established that two genes were needed for accurate assessment of gene expression. Following this, we found that horA expression was induced for Pc344NR, but not for Lb464, during growth in beer. Instead, horC expression was dramatically increased in Lb464 when growing in beer, whereas no change was detected for the other putative beer-spoilage-related genes. This indicates that HorC may be one of the principle mediators enabling growth of Lb464 in beer, whereas in Pc344NR, this may be attributable to HorA. These findings not only reveal that Lb464 and Pc344NR are unique in their beer-specific genetic expression profile but also indicate that a range of genetic specialization exists among beer-spoilage bacteria.

  13. The genome sequence of E. coli W (ATCC 9637: comparative genome analysis and an improved genome-scale reconstruction of E. coli

    Directory of Open Access Journals (Sweden)

    Lee Sang

    2011-01-01

    Full Text Available Abstract Background Escherichia coli is a model prokaryote, an important pathogen, and a key organism for industrial biotechnology. E. coli W (ATCC 9637, one of four strains designated as safe for laboratory purposes, has not been sequenced. E. coli W is a fast-growing strain and is the only safe strain that can utilize sucrose as a carbon source. Lifecycle analysis has demonstrated that sucrose from sugarcane is a preferred carbon source for industrial bioprocesses. Results We have sequenced and annotated the genome of E. coli W. The chromosome is 4,900,968 bp and encodes 4,764 ORFs. Two plasmids, pRK1 (102,536 bp and pRK2 (5,360 bp, are also present. W has unique features relative to other sequenced laboratory strains (K-12, B and Crooks: it has a larger genome and belongs to phylogroup B1 rather than A. W also grows on a much broader range of carbon sources than does K-12. A genome-scale reconstruction was developed and validated in order to interrogate metabolic properties. Conclusions The genome of W is more similar to commensal and pathogenic B1 strains than phylogroup A strains, and therefore has greater utility for comparative analyses with these strains. W should therefore be the strain of choice, or 'type strain' for group B1 comparative analyses. The genome annotation and tools created here are expected to allow further utilization and development of E. coli W as an industrial organism for sucrose-based bioprocesses. Refinements in our E. coli metabolic reconstruction allow it to more accurately define E. coli metabolism relative to previous models.

  14. The phytohormone ethylene enhances bacterial cellulose production, regulates CRP/FNRKx transcription and causes differential gene expression within the cellulose synthesis operon of Komagataeibacter (Gluconacetobacter xylinus ATCC 53582

    Directory of Open Access Journals (Sweden)

    Richard Vincent Augimeri

    2015-12-01

    Full Text Available Komagataeibacter (formerly Gluconacetobacter xylinus ATCC 53582 is a plant-associated model organism for bacterial cellulose (BC biosynthesis. This bacterium inhabits the carposphere where it interacts with fruit through the bi-directional transfer of phytohormones. The majority of research regarding K. xylinus has been focused on identifying and characterizing structural and regulatory factors that control BC biosynthesis, but its ecophysiology has been generally overlooked. Ethylene is a phytohormone that regulates plant development in a variety of ways, but is most commonly known for its positive role on fruit ripening. In this study, we utilized ethephon (2-chloroethylphosphonic acid to produce in situ ethylene to investigate the effects of this phytohormone on BC production and the expression of genes known to be involved in K. xylinus BC biosynthesis (bcsA, bcsB, bcsC, bcsD, cmcAx, ccpAx and bglAx. Using pellicle assays and reverse transcription quantitative polymerase chain reaction (RT-qPCR, we demonstrate that ethephon-derived ethylene enhances BC directly in K. xylinus by up-regulating the expression of bcsA and bcsB, and indirectly though the up-regulation of cmcAx, ccpAx and bglAx. We confirm that IAA directly decreases BC biosynthesis by showing that IAA down-regulates bcsA expression. Similarly, we confirm that ABA indirectly influences BC biosynthesis by showing it does not affect the expression of bcs operon genes. In addition, we are the first to report the ethylene and indole-3-acetic acid (IAA induced differential expression of genes within the bacterial cellulose synthesis (bcs operon. Using bioinformatics we have identified a novel phytohormone-regulated CRP/FNRKx transcription factor and provide evidence that it influences BC biosynthesis in K. xylinus. Lastly, utilizing current and previous data, we propose a model for the phytohormone-mediated fruit-bacteria interactions that K. xylinus experiences in nature.

  15. The Phytohormone Ethylene Enhances Cellulose Production, Regulates CRP/FNRKx Transcription and Causes Differential Gene Expression within the Bacterial Cellulose Synthesis Operon of Komagataeibacter (Gluconacetobacter) xylinus ATCC 53582.

    Science.gov (United States)

    Augimeri, Richard V; Strap, Janice L

    2015-01-01

    Komagataeibacter (formerly Gluconacetobacter) xylinus ATCC 53582 is a plant-associated model organism for bacterial cellulose (BC) biosynthesis. This bacterium inhabits the carposphere where it interacts with fruit through the bi-directional transfer of phytohormones. The majority of research regarding K. xylinus has been focused on identifying and characterizing structural and regulatory factors that control BC biosynthesis, but its ecophysiology has been generally overlooked. Ethylene is a phytohormone that regulates plant development in a variety of ways, but is most commonly known for its positive role on fruit ripening. In this study, we utilized ethephon (2-chloroethylphosphonic acid) to produce in situ ethylene to investigate the effects of this phytohormone on BC production and the expression of genes known to be involved in K. xylinus BC biosynthesis (bcsA, bcsB, bcsC, bcsD, cmcAx, ccpAx and bglAx). Using pellicle assays and reverse transcription quantitative polymerase chain reaction (RT-qPCR), we demonstrate that ethephon-derived ethylene enhances BC directly in K. xylinus by up-regulating the expression of bcsA and bcsB, and indirectly though the up-regulation of cmcAx, ccpAx, and bglAx. We confirm that IAA directly decreases BC biosynthesis by showing that IAA down-regulates bcsA expression. Similarly, we confirm that ABA indirectly influences BC biosynthesis by showing it does not affect the expression of bcs operon genes. In addition, we are the first to report the ethylene and indole-3-acetic acid (IAA) induced differential expression of genes within the bacterial cellulose synthesis (bcs) operon. Using bioinformatics we have identified a novel phytohormone-regulated CRP/FNRKx transcription factor and provide evidence that it influences BC biosynthesis in K. xylinus. Lastly, utilizing current and previous data, we propose a model for the phytohormone-mediated fruit-bacteria interactions that K. xylinus experiences in nature.

  16. Influence of sub-inhibitory antibiotics and flow condition on Staphylococcus aureus ATCC 6538 biofilm development and biofilm growth rate: BioTimer assay as a study model.

    Science.gov (United States)

    Berlutti, Francesca; Frioni, Alessandra; Natalizi, Tiziana; Pantanella, Fabrizio; Valenti, Piera

    2014-11-01

    Staphylococcus biofilm exhibits high antibiotic resistance and therapeutic doses of antibiotics are often sub-inhibitory. Whereas data are available on the effect of sub-inhibitory antibiotics on matrix formation, little is known on their influence on biofilm population. Here, using BioTimer Assay (BTA), a method developed to quantify biofilm population, the influence of sub-inhibitory gentamicin, ofloxacin and azithromycin on Staphylococcus aureus ATCC 6538 biofilm population in flow with respect to static condition was assessed. Antibiotics and flow condition increased biofilm population even if at different extent, depending on the antibiotic molecule. The greatest bacterial population was found in biofilm developed under flow condition in the presence of azithromycin. A significant increase in biofilm matrix was recorded for biofilm developed in the presence of antibiotics in flow with respect to static condition. The growth rates (GRs) of 24-h biofilm developed under the influence of antibiotics and flow condition were also evaluated using BTA and a specific mathematical model. Antibiotics and flow condition affected the GRs of 24-h biofilm even if at different extent. The lowest GR value was recorded for biofilm developed under flow condition in the presence of ofloxacin. Although further studies are needed, our data indicate that antibiotics and flow condition influenced biofilm development by increasing both bacterial population and matrix formation and affected the GRs of the developed biofilm. To the best of our knowledge, BTA is unique in allowing the calculation of the GRs of biofilm and it may be considered to be a useful study model to evaluate the activity of antibiofilm molecules. PMID:24865865

  17. Daya Antibakteri Filtrat Asam Laktat dan Bakteriosin Lactobacillus bulgaricus KS1 dalam Menghambat Pertumbuhan Klebsiella pneumoniae Strain ATCC 700603, CT1538, dan S941

    Directory of Open Access Journals (Sweden)

    Prima Nanda Fauziah

    2015-03-01

    Full Text Available Lactobacillus bulgaricus produces lactic acid and bacteriocin which have been reported to have various pharmacologic properties, including their role an antibacterial agent. Klebsiella pneumoniae, as an agent of pneumonia, remains a public health problem in tropical countries. This study was aimed to observe the antibacterial activities of lactic acid filtrate and bacteriocins of L. bulgaricus toward againsts K. pneumoniae strains by in vitro experiment. The experiment took place in Microbiology Laboratory, Teaching Hospital, Padjadjaran University, Bandung, August–October 2012. In vitro laboratory analytic study has been conducted on lactic acid filtrate and bacteriocins of L. bulgaricus against the K. pneumoniae strains. The study used agar pour plate and agar disk diffusion method and analyzed by ANAVA followed by Duncan’s multiple range test (DMRT. The 30% lactic acid filtrate and 20% bacteriocins filtrate concentrations of L. bulgaricus showed bactericidal characteristics againts the growth of K. pneumoniae strains. Greater concentration of lactic acid filtrate and bacteriocins of L. bulgaricus led toincreasing effect of growth inhibition zones of K. pneumoniae strains. Statistical analysis of variance (ANOVA showed that the greatest concentration effect of L. bulgaricus filtratefor inhibiting K. pneumoniae strains was achieved in 90% lactic acid filtrate concentration treatment, whereas the greatest inhibition zones for K. pneumoniae ATCC 700603 was obtaubed in 90% bacteriocins filtrate concentration, amounting 16.667 mm. In conclusion, lactic acid filtrate and bacteriocins L. bulgaricus have antibacterial effects on K. pneumoniae. The level of antibacterial effect of L. bulgaricus against the growth of K. pneumoniae strains depends on the type of filtrate, L. bulgaricus filtrate concentration, and K. pneumoniae strain.

  18. Applications of the Box-Wilson design model for bio-hydrogen production using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564).

    Science.gov (United States)

    Alalayah, W M; Kalil, M S; Kadhum, A A H; Jahim, J; Zaharim, A; Alauj, N M; El-Shafie, A

    2010-07-15

    Box-Wilson design (BWD) model was applied to determine the optimum values of influencing parameters in anaerobic fermentation to produce hydrogen using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). The main focus of the study was to find the optimal relationship between the hydrogen yield and three variables including initial substrate concentration, initial medium pH and reaction temperature. Microbial growth kinetic parameters for hydrogen production under anaerobic conditions were determined using the Monod model with incorporation of a substrate inhibition term. The values of micro(max) (maximum specific growth rate) and K, (saturation constant) were 0.398 h(-1) and 5.509 g L(-1), respectively, using glucose as the substrate. The experimental substrate and biomass-concentration profiles were in good agreement with those obtained by the kinetic-model predictions. By varying the conditions of the initial substrate concentration (1-40 g L(-1)), reaction temperature (25-40 degrees C) and initial medium pH (4-8), the model predicted a maximum hydrogen yield of 3.24 mol H2 (mol glucose)(-1). The experimental data collected utilising this design was successfully fitted to a second-order polynomial model. An optimum operating condition of 10 g L(-1) initial substrate concentration, 37 degrees C reaction temperature and 6.0 +/- 0.2 initial medium pH gave 80% of the predicted maximum yield of hydrogen where as the experimental yield obtained in this study was 77.75% exhibiting a close accuracy between estimated and experimental values. This is the first report to predict bio-hydrogen yield by applying Box-Wilson Design in anaerobic fermentation while optimizing the effects of environmental factors prevailing there by investigating the effects of environmental factors.

  19. An investigation of agitation speed as a factor affecting the quantity and monomer distribution of alginate from Azotobacter vinelandii ATCC(®) 9046.

    Science.gov (United States)

    Kıvılcımdan Moral, C; Sanin, F D

    2012-03-01

    Alginate is a copolymer of β-D: -mannuronic and α-L: -guluronic acids. Distribution of these monomers in the alginate structure is one of the important characteristics that affect the commercial value of the polymer. In the present work, the effect of agitation speed in the range of 200-700 rpm on alginate production by Azotobacter vinelandii ATCC(®) 9046 was investigated at a dissolved oxygen tension of 5% of air saturation. Experiments were conducted in a fermentor operated in batch mode for 72 h while the production of biomass and alginate, the consumption of substrate and the change in culture broth viscosity and monomer distribution of the polymer were monitored. Results showed that the growth rate of the bacteria increased from 0.165 to 0.239 h(-1) by the increase of mixing speed from 200 to 400 rpm. On the other hand, alginate production was found to be the most efficient at 400 rpm with the highest value of 4.51 g/l achieved at the end of fermentation. The viscosity of culture broth showed similar trends to alginate production. Viscosity was recorded as 24.61 cP at 400 rpm while it was only 4.26 cP at 700 rpm. The MM- and GG-block contents were almost equal in most of the culture times at 400 rpm. On the other hand, GG-blocks dominated at both low and high mixing speeds. Knowing that GG-blocks make rigid and protective gels with divalent cations, due to the higher GG-block content, the gel formation potential is higher at 200 rpm as well at 700 rpm, which might originate from the unfavorable environmental conditions that the bacteria were exposed to. PMID:22009058

  20. Cloning, sequencing and partial characterisation of sorbitol transporter (srlT) gene encoding phosphotransferase system, glucitol/sorbitol-specific IIBC components of Erwinia herbicola ATCC 21998.

    Science.gov (United States)

    Qazi, P H; Johri, S; Verma, V; Khan, L; Qazi, G N

    2004-09-01

    A DNA fragment of approximately 1500 bp, harbouring the sorbitol transport gene (srlT), was amplified from the chromosomal DNA of Erwinia herbicola ATCC 21998 by PCR and cloned in Escherichia coli JM109. Degenerate oligonucleotide primers used were designed based on the conserved regions in the gene sequences within the gut operon of E. coli (Gene Bank accession no. J02708) and the srl operon of Erwinia amylovora (Gene Bank accession no. Y14603). The cloned DNA fragment was sequenced and found to contain an open reading frame of 1473 nucleotides coding for a protein of 491 amino acids, corresponding to a mass of 52410 Da. The nucleotide sequence of this ORF was highly homologous to that of the gutA gene of Escherichia coli gut operon, the srlE gene of Shigella flexrni and the sorbitol transporter gene sequence of Escherichia coli K12 (Gene Bank accession nos. J02708, AE016987 and D90892 respectively). The protein sequence showed significant homology to that of the phosphotransferase system i.e. the glucitol/sorbitol-specific IIBC components of Escherichia coli and Erwinia amylovora (P56580, O32522). The cloned DNA fragment was introduced into a pRA90 vector and the recombinant was used for developing srlT mutants of Erwinia herbicola, by homologous recombination. Mutants obtained were unable to grow on minimal medium with sorbitol. The insertion of the pRA90 vector inside the srlT gene sequence of the mutants was confirmed by DNA-DNA hybridisation. PMID:15560368

  1. Effects of Bacillus subtilis 'PB6' (ATCC - PTA 6737 on Clostridium difficile Associated Diarrhea (CDAD and Inflammatory Bowel Disease (IBD in Animal Models

    Directory of Open Access Journals (Sweden)

    Eric Peys

    2007-01-01

    Full Text Available The administration of probiotic bacteria is emerging as a potential means of preventing the onset or recurrence of Clostridium difficile associated diarrhea (CDAD and of attenuating inflammatory activity and preventing relapses in inflammatory bowel disease (IBD. We evaluated the efficacy of Bacillus subtilis ‘PB6’ (ATCC – PTA 6737 in a hamster model of antibiotic-induced CDAD and in a rat model of IBD. CDAD was induced in male Golden Syrian hamsters using C. difficile and clindamycin. These hamsters received either nothing or, by gavage, vancomycin (5 days or PB6 (low, middle and high dose, 6 days. Diarrhea, body weight loss and mortality were observed in all groups in which CDAD was induced. Intensity of diarrhea and body weight loss was least in the groups treated with vancomycin or with the highest dose of PB6. At the end of the treatment period, vancomycin and the highest dose of PB6 were equally efficient in preventing mortality in this hamster model of CDAD. No adverse effects of PB6 treatment were observed in healthy animals. In male Wistar rats, colitis was induced using a single intrarectal administration of 2,4,6-trinitrobenzenesulfonic acid (TNBS. Treatments consisted of PB6 (low, middle and high dose, Saccharomyces boulardii, mesalazine, infliximab, or no treatment. A possible benefit of the prophylactic use of PB6 was also tested. At the end of the treatment period significant differences in body weight gain, in colon inflammatory edema and in gross morphology of the colon intestinal lining were observed between groups. The groups treated with high dose PB6 could not be distincted from the colitis-free negative control group nor from the group treated with mesalazine. The data presented are suggestive of possible therapeutic effectiveness of PB6 in CDAD and IBD in humans.

  2. The Phytohormone Ethylene Enhances Cellulose Production, Regulates CRP/FNRKx Transcription and Causes Differential Gene Expression within the Bacterial Cellulose Synthesis Operon of Komagataeibacter (Gluconacetobacter) xylinus ATCC 53582.

    Science.gov (United States)

    Augimeri, Richard V; Strap, Janice L

    2015-01-01

    Komagataeibacter (formerly Gluconacetobacter) xylinus ATCC 53582 is a plant-associated model organism for bacterial cellulose (BC) biosynthesis. This bacterium inhabits the carposphere where it interacts with fruit through the bi-directional transfer of phytohormones. The majority of research regarding K. xylinus has been focused on identifying and characterizing structural and regulatory factors that control BC biosynthesis, but its ecophysiology has been generally overlooked. Ethylene is a phytohormone that regulates plant development in a variety of ways, but is most commonly known for its positive role on fruit ripening. In this study, we utilized ethephon (2-chloroethylphosphonic acid) to produce in situ ethylene to investigate the effects of this phytohormone on BC production and the expression of genes known to be involved in K. xylinus BC biosynthesis (bcsA, bcsB, bcsC, bcsD, cmcAx, ccpAx and bglAx). Using pellicle assays and reverse transcription quantitative polymerase chain reaction (RT-qPCR), we demonstrate that ethephon-derived ethylene enhances BC directly in K. xylinus by up-regulating the expression of bcsA and bcsB, and indirectly though the up-regulation of cmcAx, ccpAx, and bglAx. We confirm that IAA directly decreases BC biosynthesis by showing that IAA down-regulates bcsA expression. Similarly, we confirm that ABA indirectly influences BC biosynthesis by showing it does not affect the expression of bcs operon genes. In addition, we are the first to report the ethylene and indole-3-acetic acid (IAA) induced differential expression of genes within the bacterial cellulose synthesis (bcs) operon. Using bioinformatics we have identified a novel phytohormone-regulated CRP/FNRKx transcription factor and provide evidence that it influences BC biosynthesis in K. xylinus. Lastly, utilizing current and previous data, we propose a model for the phytohormone-mediated fruit-bacteria interactions that K. xylinus experiences in nature. PMID:26733991

  3. Characterization of the extended-spectrum beta-lactamase reference strain, Klebsiella pneumoniae K6 (ATCC 700603), which produces the novel enzyme SHV-18.

    Science.gov (United States)

    Rasheed, J K; Anderson, G J; Yigit, H; Queenan, A M; Doménech-Sánchez, A; Swenson, J M; Biddle, J W; Ferraro, M J; Jacoby, G A; Tenover, F C

    2000-09-01

    Klebsiella pneumoniae K6 (ATCC 700603), a clinical isolate, is resistant to ceftazidime and other oxyimino-beta-lactams. A consistent reduction in the MICs of oxyimino-beta-lactams by at least 3 twofold dilutions in the presence of clavulanic acid confirmed the utility of K. pneumoniae K6 as a quality control strain for extended-spectrum beta-lactamase (ESBL) detection. Isoelectric-focusing analysis of crude lysates of K6 demonstrated a single beta-lactamase with a pI of 7.8 and a substrate profile showing preferential hydrolysis of cefotaxime compared to ceftazidime. PCR analysis of total bacterial DNA from K6 identified the presence of a bla(SHV) gene. K6 contained two large plasmids with molecular sizes of approximately 160 and 80 kb. Hybridization of plasmid DNA with a bla(SHV)-specific probe indicated that a bla(SHV) gene was encoded on the 80-kb plasmid, which was shown to transfer resistance to ceftazidime in conjugal mating experiments with Escherichia coli HB101. DNA sequencing of this bla(SHV)-related gene revealed that it differs from bla(SHV-1) at nine nucleotides, five of which resulted in amino acid substitutions: Ile to Phe at position 8, Arg to Ser at position 43, Gly to Ala at position 238, and Glu to Lys at position 240. In addition to the production of this novel ESBL, designated SHV-18, analysis of the outer membrane proteins of K6 revealed the loss of the OmpK35 and OmpK37 porins. PMID:10952583

  4. Effect of Shuanghuanglian Combined with Levofloxacin on Antibiotic Resistance of Staphylococcus Aureus ATCC29213 in Rabbit Tissue Cage Infection Model%双黄连联用左氧氟沙星对兔组织笼感染模型中金黄色葡萄球菌ATCC29213耐药性的影响

    Institute of Scientific and Technical Information of China (English)

    王国俊; 叶云; 冯碧敏; 李虹

    2014-01-01

    Objective To explore effects of shuanghuanglian combined with levofloxacin on antibiotic resistance of Staphylococcus Aureus ATCC29213 to levofloxacin. Methods Tissue cage infection model with Staphylococcus aureus was established in rabbits, and the infected animals were given with levofloxacin alone ( group A ) or in combination with shuanghuanglian ( group B) for 5 days respectively. Steady-state concentration of levofloxacin in tissue cage, bacteria recovery and bacterial resistance in tissue cage infection model were studied. Results Steady-state concentration of levofloxacin in tissue cage was not significantly different between group A and group B. The recovery rate of bacteria was significantly lower in group B than in group A (20. 0% vs. 100. 0%). The minimum inhibitory concentration (MIC) was lower in group B than in group A. Conclusion Shuanghuanglian combined with levofloxacin is helpful to reduce antibiotic resistance of Staphylococcus aureus to levofloxacin, indicating that some Chinese traditional medicine combined with antibiotics can reduce antibiotic resistance.%目的探讨双黄连联合左氧氟沙星对兔组织笼感染模型中金黄色葡萄球菌ATCC29213对左氧氟沙星的耐药性的影响。方法建立兔组织笼感染模型,分别用左氧氟沙星单独处理(A组)及双黄连联合左氧氟沙星处理(B组)5 d,探讨兔组织笼感染模型中左氧氟沙星在组织液中稳态浓度及经处理后兔组织笼中细菌恢复生长的情况和细菌耐药性变化。结果两组左氧氟沙星在组织液中浓度差异无统计学意义。 B组兔组织笼内细菌恢复生长发生率(20%)明显低于A组(100.0%),而左氧氟沙星对B组兔组织笼内细菌的最低抑菌浓度(MIC)值明显低于A组。结论双黄连与左氧氟沙星联合治疗有助于减少金黄色葡萄球菌对左氧氟沙星的耐药现象,提示中药联合抗菌药物可降低细菌对抗菌药物的耐药性。

  5. Construction and General Characteristics of the plcR Mutation in Bacillus cereus ATCC14579%蜡样芽孢杆菌ATCC14579毒力基因plcR缺失株的构建及其一般特性

    Institute of Scientific and Technical Information of China (English)

    吴威; 郑经成; 闫广谋; 王思洮; 盛相鹏; 胡丹; 张阳阳; 韩文瑜

    2010-01-01

    蜡样芽孢杆菌是土壤中的优势菌,具有作为益生菌的潜在能力,同时它也是条件致病菌,能引起食物中毒等.蜡样芽孢杆菌的多种毒力因子受到多效性调控子(pleiotropic regulator,plcR)的调控,在其条件致病性作用中起着重要作用.真养产碱杆菌JMP34(Alcaligelles eutrophus)质粒上的2,4-二氯苯氧乙酸(2,4-dichlorophenoxyacetic acid,2,4-D)单加氧酶(tfdA)基因可以降解2,4-D.本研究利用同源重组技术使tfdA基因置换掉蜡样芽孢杆菌的plcR基因,构建了蜡样芽孢杆菌ATCC14579突变株B.cereus ΔplcRΩtf-dA,并对其毒性、一般生理生化特性进行分析.研究结果表明,突变株B.cereus ΔplcRΩtfdA的毒性显著减弱;生理生化实验结果显示突变株与野生株并没有明显区别,且突变株并没有表现出tfdA酶活性.所有的结果表明plcR控制着蜡样芽孢杆菌ATCC14579的致病性,同时剔除plcR并不破坏其酶系统.本研究为今后构建蜡样芽孢杆菌工程菌提供了新的思路和依据.

  6. Clostridium scindens ATCC 35704中的D-塔格糖-3-差向异构酶的克隆表达、纯化及活性研究%Cloning, Expression, Purification and Characterization of D-Tagatose-3-Epimerase from Clostridium scindens ATCC 35704

    Institute of Scientific and Technical Information of China (English)

    邢庆超; 沐万孟; 江波; 周榴明; 张涛

    2011-01-01

    D-tagatose-3-epimerase (DTE) catalyzes the epimerization of various ketohexoses at C-3 position, and commonly catalyzes the reversible interconversion of D-fructose to D-psicose, which is a novel useful low-calorie sweetener with many beneficial effects. A new gene, encoding the hypothetical protein with locus ZP 02432283 from the Clostridium scindens ATCC 35704 was cloned and over-expressed in E. coli. BL21 (DE3). The recombinant protein was purified to electrophoretical homogeneity by affinity chromatography using Chelating Sepharose Fast Flow resin and analyzed by SDS-PAGE, showing approximately 31 ku characteristic protein. The recombinant enzyme activity was determined by measuring D-sorbose formed using D-tagatose and D-psicose formed using D-fructose as a substrate, the bioconversion rate reached 8.6% and 27.9% , respectively.%D-阿洛酮糖作为一种新型低热量功能性甜味剂,可以通过D-塔格糖-3-差向异构酶家族,以D-果糖为底物C-3位异构化得到。一个新的来源于微生物Clostridium scindens ATCC 35704中ZP 0243228的D-塔格糖-3-差向异构酶基因(CS-DTE),通过克隆并成功导入E.coli BL21(DE3)中,构建了基因重组菌,诱导目的重组基因过量表达;经亲和层析纯化的重组蛋白样品进行SDS—PAGE分析,在约31ku处出现显著的特征蛋白条带;通过对其活性检测表明,该重组酶分别以D-塔格糖和D-果糖为底物,可以生成D-山梨糖和D-阿洛酮糖,转化率分别为8.6%和27.9%。

  7. Conserved genes in a path from commensalism to pathogenicity: comparative phylogenetic profiles of Staphylococcus epidermidis RP62A and ATCC12228

    Directory of Open Access Journals (Sweden)

    Jia PeiLin

    2006-05-01

    Full Text Available Abstract Background Staphylococcus epidermidis, long regarded as an innocuous commensal bacterium of the human skin, is the most frequent cause of nosocomial infections associated with implanted medical devices. This conditional pathogen provides a model of choice to study genome landmarks correlated with the transition between commensalism and pathogenicity. Traditional investigations stress differences in gene content. We focused on conserved genes that have accumulated small mutation differences during the transition. Results A comparison of strain ATCC12228, a non-biofilm forming, non-infection associated strain and strain RP62A, a methicillin-resistant biofilm clinical isolate, revealed consistent variation, mostly single-nucleotide polymorphisms (SNPs, in orthologous genes in addition to the previously investigated global changes in gene clusters. This polymorphism, scattered throughout the genome, may reveal genes that contribute to adaptation of the bacteria to different environmental stimuli, allowing them to shift from commensalism to pathogenicity. SNPs were detected in 931 pairs of orthologs with identical gene length, accounting for approximately 45% of the total pairs of orthologs. Assuming that non-synonymous mutations would mark recent evolution, and hence be associated to the onset of the pathogenic process, analysis of ratios of non-synonymous SNPs vs synonymous SNPs suggested hypotheses about possible pathogenicity determinants. The N/S ratios for virulence factors and surface proteins differed significantly from that of average SNPs. Of those gene pairs, 40 showed a disproportionate distribution of dN vs dS. Among those, the presence of the gene encoding methionine sulfoxide reductase suggested a possible involvement of reactive oxygen species. This led us to uncover that the infection associated strain was significantly more resistant to hydrogen peroxide and paraquat than the environmental strain. Some 16 genes of the list

  8. Helicobacter pylori ATCC 43629/NCTC 11639 outer membrane vesicles (OMVs from biofilm and planktonic phase associated with extracellular DNA (eDNA.

    Directory of Open Access Journals (Sweden)

    Rossella eGrande

    2015-12-01

    Full Text Available Helicobacter pylori persistence is associated to its capability of developing biofilms as a response to environmental stress and changes. Extracellular DNA (eDNA is a component of H. pylori biofilm matrix but the lack of DNase I activity supports the hypothesis that eDNA might be protected by other extracellular polymeric substances and/or Outer Membrane Vesicles (OMVs, which bleb from the bacteria surface during growth. The aim of the present study was to both identify the eDNA presence on OMVs segregated from H. pylori biofilm (bOMVs and its planktonic phase (pOMVs and to characterize the physical-chemical properties of bOMVs and pOMVs. The presence of eDNA in bOMVs and pOMVs was carried out using a DNase I-gold complex and Transmission Electron Microscope analysis (TEM. bOMVs and pOMVs were further isolated and physical-chemical characterized using the dynamic light scattering (DLS analysis. The eDNA associated to OMVs was detected and quantified by using PicoGreen assay and spectrophotometer, while its extraction was performed through a DNA Kit. The TEM images showed that the eDNA was mainly isolated and identified on OMVs-membrane surface; while the PicoGreen staining showed a 4-fold increase of dsDNA in bOMVs compared to pOMVs. The eDNA extracted from OMVs was visualized by using gel electrophoresis. The DLS analysis demonstrated that H. pylori generate vesicles, both in its planktonic and biofilm phenotypes, with sizes in the nanometer scales and a broad size distribution. The DLS aggregation study of H. pylori OMVs demonstrated that eDNA may play a role in the OMVs aggregation, particularly for biofilm phenotype. The eDNA associated with vesicle membrane can affect the DNase I activity on H. pylori biofilms. OMVs derived from H. pylori ATCC 43629/NCTC 11639, particularly its biofilm phenotype, may play a structural role by preventing eDNA degradation by nucleases and provide a bridging function between eDNA strands.

  9. Initial reaction(s) in biotransformation of CL-20 is catalyzed by salicylate 1-monooxygenase from Pseudomonas sp. strain ATCC 29352.

    Science.gov (United States)

    Bhushan, Bharat; Halasz, Annamaria; Spain, Jim C; Hawari, Jalal

    2004-07-01

    CL-20 (2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane) (C(6)H(6)N(12)O(12)), a future-generation high-energy explosive, is biodegradable by Pseudomonas sp. strain FA1 and Agrobacterium sp. strain JS71; however, the nature of the enzyme(s) involved in the process was not understood. In the present study, salicylate 1-monooxygenase, a flavin adenine dinucleotide (FAD)-containing purified enzyme from Pseudomonas sp. strain ATCC 29352, biotransformed CL-20 at rates of 0.256 +/- 0.011 and 0.043 +/- 0.003 nmol min(-1) mg of protein(-1) under anaerobic and aerobic conditions, respectively. The disappearance of CL-20 was accompanied by the release of nitrite ions. Using liquid chromatography/mass spectrometry in the negative electrospray ionization mode, we detected a metabolite with a deprotonated mass ion [M - H](-) at 345 Da, corresponding to an empirical formula of C(6)H(6)N(10)O(8), produced as a result of two sequential N denitration steps on the CL- 20 molecule. We also detected two isomeric metabolites with [M - H](-) at 381 Da corresponding to an empirical formula of C(6)H(10)N(10)O(10). The latter was a hydrated product of the metabolite C(6)H(6)N(10)O(8) with addition of two H(2)O molecules, as confirmed by tests using (18)O-labeled water. The product stoichiometry showed that each reacted CL-20 molecule produced about 1.7 nitrite ions, 3.2 molecules of nitrous oxide, 1.5 molecules of formic acid, and 0.6 ammonium ion. Diphenyliodonium-mediated inhibition of salicylate 1-monooxygenase and a comparative study between native, deflavo, and reconstituted enzyme(s) showed that FAD site of the enzyme was involved in the biotransformation of CL-20 catalyzed by salicylate 1-monooxygenase. The data suggested that salicylate 1-monooxygenase catalyzed two oxygen-sensitive single-electron transfer steps necessary to release two nitrite ions from CL-20 and that this was followed by the secondary decomposition of this energetic chemical. PMID:15240281

  10. Validation of Baking To Control Salmonella Serovars in Hamburger Bun Manufacturing, and Evaluation of Enterococcus faecium ATCC 8459 and Saccharomyces cerevisiae as Nonpathogenic Surrogate Indicators.

    Science.gov (United States)

    Channaiah, Lakshmikantha H; Holmgren, Elizabeth S; Michael, Minto; Sevart, Nicholas J; Milke, Donka; Schwan, Carla L; Krug, Matthew; Wilder, Amanda; Phebus, Randall K; Thippareddi, Harshavardhan; Milliken, George

    2016-04-01

    This study was conducted to validate a simulated commercial baking process for hamburger buns to destroy Salmonella serovars and to determine the appropriateness of using nonpathogenic surrogates (Enterococcus faecium ATCC 8459 or Saccharomyces cerevisiae) for in-plant process validation studies. Wheat flour was inoculated (∼6 log CFU/g) with three Salmonella serovars (Typhimurium, Newport, or Senftenberg 775W) or with E. faecium. Dough was formed, proofed, and baked to mimic commercial manufacturing conditions. Buns were baked for up to 13 min in a conventional oven (218.3°C), with internal crumb temperature increasing to ∼100°C during the first 8 min of baking and remaining at this temperature until removal from the oven. Salmonella and E. faecium populations were undetectable by enrichment (>6-log CFU/g reductions) after 9.0 and 11.5 min of baking, respectively, and ≥5-log-cycle reductions were achieved by 6.0 and 7.75 min, respectively. D-values of Salmonella (three-serovar cocktail) and E. faecium 8459 in dough were 28.64 and 133.33, 7.61 and 55.67, and 3.14 and 14.72 min at 55, 58, and 61°C, respectively, whereas D-values of S. cerevisiae were 18.73, 5.67, and 1.03 min at 52, 55, and 58°C, respectivly. The z-values of Salmonella, E. faecium, and S. cerevisiae were 6.58, 6.25, and 4.74°C, respectively. A high level of thermal lethality was observed for baking of typical hamburger bun dough, resulting in rapid elimination of high levels of the three-strain Salmonella cocktail; however, the lethality and microbial destruction kinetics should not be extrapolated to other bakery products without further research. E. faecium demonstrated greater thermal resistance compared with Salmonella during bun baking and could serve as a conservative surrogate to validate thermal process lethality in commercial bun baking operations. Low thermal tolerance of S. cerevisiae relative to Salmonella serovars limits its usefulness as a surrogate for process validations

  11. Complete genome sequence and lifestyle of black-pigmented Corynebacterium aurimucosum ATCC 700975 (formerly C. nigricans CN-1 isolated from a vaginal swab of a woman with spontaneous abortion

    Directory of Open Access Journals (Sweden)

    Gartemann Karl-Heinz

    2010-02-01

    Full Text Available Abstract Background Corynebacterium aurimucosum is a slightly yellowish, non-lipophilic, facultative anaerobic member of the genus Corynebacterium and predominantly isolated from human clinical specimens. Unusual black-pigmented variants of C. aurimucosum (originally named as C. nigricans continue to be recovered from the female urogenital tract and they are associated with complications during pregnancy. C. aurimucosum ATCC 700975 (C. nigricans CN-1 was originally isolated from a vaginal swab of a 34-year-old woman who experienced a spontaneous abortion during month six of pregnancy. For a better understanding of the physiology and lifestyle of this potential urogenital pathogen, the complete genome sequence of C. aurimucosum ATCC 700975 was determined. Results Sequencing and assembly of the C. aurimucosum ATCC 700975 genome yielded a circular chromosome of 2,790,189 bp in size and the 29,037-bp plasmid pET44827. Specific gene sets associated with the central metabolism of C. aurimucosum apparently provide enhanced metabolic flexibility and adaptability in aerobic, anaerobic and low-pH environments, including gene clusters for the uptake and degradation of aromatic amines, L-histidine and L-tartrate as well as a gene region for the formation of selenocysteine and its incorporation into formate dehydrogenase. Plasmid pET44827 codes for a non-ribosomal peptide synthetase that plays the pivotal role in the synthesis of the characteristic black pigment of C. aurimucosum ATCC 700975. Conclusions The data obtained by the genome project suggest that C. aurimucosum could be both a resident of the human gut and possibly a pathogen in the female genital tract causing complications during pregnancy. Since hitherto all black-pigmented C. aurimucosum strains have been recovered from female genital source, biosynthesis of the pigment is apparently required for colonization by protecting the bacterial cells against the high hydrogen peroxide concentration in

  12. Conditions for Protoplast Preparation and Regeneration by Sphingomonas sp. ATCC 31555%韦兰胶合成菌的原生质体制备与再生研究

    Institute of Scientific and Technical Information of China (English)

    周明明; 李晓雁; 任梦楠; 程珂萌; 黄海东

    2016-01-01

    旨在研究 EDTA 预处理、菌龄、酶解浓度、酶解温度及酶解时间对 Sphingomonas sp. ATCC 31555原生质体制备与再生的影响。结果表明,菌龄10 h,12 mmol/L EDTA 预处理,溶菌酶浓度45μg/mL、酶解温度28℃、酶解时间25 min。得到原生质体的形成率达97.3%,再生率达33.9%。研究结果为菌株进行原生质体融合、基因组改组等选育优良菌种提供参考。%The effects of EDTA pretreatment,cell age and the concentration,temperature and work time of enzymolysis on protoplast formation and regeneration by Sphingomonas sp. ATCC 31555 were studied. The results showed that the formation rate and regeneration rate reached 97.3% and 33.9% respectively,when the cells cultivated 10 h,pretreated by 12 mmol/L EDTA,using 45 μg/mL of lysozyme, enzymolysis at 28℃ for 25 min. The result of this paper provides a reference for breeding fine microbial strains by protoplast fusion and genome shuffling.

  13. Nitric oxide production in celomocytes of the earthworm Eisenia hortensis following bacterial challenge

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    SR Cook

    2015-02-01

    Full Text Available In this in vitro investigation, nitric oxide (NO production was induced within celomocytes of the earthworm Eisenia hortensis following microbial challenge. Celomocytes were pre-loaded with the fluorescent indicator 4-amino-5-methylamino-2’, 7’-difluorofluorescein diacetate (DAF-FM DA in order to detect the presence of intracellular nitric oxide subsequent to a 16 h incubation with chemically-fixed soil bacteria including Bacillus megaterium, Arthrobacter globiformis, Pseudomonas stutzeri, and Azotobacter chroococcum at a range of multiplicities of infection (MOIs. Flow cytometric analysis measuring increases in relative fluorescence intensity (RFI, which is directly proportional to the amount of intracellular NO produced, permitted determination of statistical significance (p < 0.05 of exposed celomocytes compared to baseline controls. Significant increases in NO were detected reproducibly in celomocytes treated with all bacterial species used. The most prominent results were observed after exposure to Gram positive B. megaterium and A. globiformis where 100 % of earthworms tested exhibited statistically significant increases of RFI at MOIs of 100:1 and 500:1, respectively. Furthermore, significant decreases in NO production in bacteria-stimulated earthworm celomocytes incubated with the NOS inhibitor aminoguanidine hydrochloride were observed. These results demonstrate microbial induction of NO synthesis in earthworms and provide evidence of an antimicrobial role of NO in the innate immune system.

  14. Arthrobacter enclensis sp. nov., isolated from sediment sample

    Digital Repository Service at National Institute of Oceanography (India)

    Dastager, S.G.; Qin, L.; Tang, S.K.; Krishnamurthi, S.; Lee, J.C.; Li, W.J.

    ,000 replicates. The DNA G+C content (mol%) was estimated accord- ing to Gonzalez and Saiz-Jimenez (2002). ΔTm values between homologous and hybrid DNA are analysed by real-time PCR (Gonzalez and Saiz-Jimenez 2005). For DNA–DNA hybridization experiments...:228–230 Gillis M, De Ley J, De Cleene M (1970) The determination of molec- ular weight of bacterial genome DNA from renaturation rates. Eur J Biochem 12:143–153 Gonzalez JM, Saiz-Jimenez C (2002) A fluorimetric method for the estimation of G+C mol% content...

  15. Beta-blockers in the environment: part II. Ecotoxicity study.

    Science.gov (United States)

    Maszkowska, Joanna; Stolte, Stefan; Kumirska, Jolanta; Łukaszewicz, Paulina; Mioduszewska, Katarzyna; Puckowski, Alan; Caban, Magda; Wagil, Marta; Stepnowski, Piotr; Białk-Bielińska, Anna

    2014-09-15

    The increasing consumption of beta-blockers (BB) has caused their presence in the environment to become more noticeable. Even though BB are safe for human and veterinary usage, ecosystems may be exposed to these substances. In this study, three selected BB: propranolol, metoprolol and nadolol were subjected to ecotoxicity study. Ecotoxicity evaluation was based on a flexible ecotoxicological test battery including organisms, representing different trophic levels and complexity: marine bacteria (Vibrio fischeri), soil/sediment bacteria (Arthrobacter globiformis), green algae (Scenedesmus vacuolatus) and duckweed (Lemna minor). All the ecotoxicological studies were supported by instrumental analysis to measure deviation between nominal and real test concentrations. Based on toxicological data from the green algae test (S. vacuolatus) propranolol and metoprolol can be considered to be harmful to aquatic organisms. However, sorption explicitly inhibits the hazardous effects of BB, therefore the risks posed by these compounds for the environment are of minor importance. PMID:24975494

  16. 基于细胞性质分析不同有机溶剂对简单节杆菌C1,2脱氢反应的影响%Study on the Effects of Different Kinds of Organic Solvents on C1,2 Dehydrogenation Reaction by the Analysis of Arthrobacter Simplex Cell Characters

    Institute of Scientific and Technical Information of China (English)

    骆健美; 宋妍; 王建锋; 宁静; 成永新; 郑宇; 申雁冰

    2012-01-01

    通过对简单节杆菌细胞性质的研究,探讨了不同有机溶剂(乙醇、DMF、DMSO、甲醇)加入后醋酸可的松生物转化反应效率产生差异的原因.采用底物转化法检测生物转化率,采用平板活菌计数法计算细胞存活率,通过糖代谢活力保留值表征细胞的生理代谢活力,用原子力显徽镜观察细胞超显微结构,以FDA法表征细胞膜通透性.在体积分数为4%的条件下,4种有机溶剂均能提高简单节杆菌生物转化醋酸可的松的初始转化速率和最终转化率,转化率从高到低依次为乙醇>DMSO>甲醇>DMF;4种溶剂对简单节杆菌存活率和糖代谢活力保留值的影响趋势基本一致,其中DMF最大,DMSO次之,乙醇和甲醇较小;4种溶剂中,甲醇对简单节杆菌细胞完整性的影响最大,乙醇次之,DMF和DMSO差别不明显;4种有机溶剂对简单节杆菌细胞膜通透性的影响大小依次为甲醇>乙醇>DMSO>DMF.上述结果表明,细胞生理代谢活力和细胞膜通透性的差异是造成转化率水平不同的重要原因.其中,细胞膜通透性的增强有利于底物分子更容易进入细胞与生物酶接触,进而提高转化率.%The cell characters were studied to analyze the reason for difference of the conversion efficiency of Arthrobacter simplex on cortisoniacetas(CA) in the presence of different kinds of organic solvents (ethanol,DMF,DMSO,methanol).The bioconversion rate was determined by the substrate conversion method,the cell survival ratio was calculated by the counting method on plates,the cell metabolic activity was assessed by the glucose metabolic activity retention,AFM was used to observe the cell ultrastructure,FDA method was adopted to determine the cell membrane permeability.Both of the initial conversion rate and the final conversion rate for the biotransformation process were increased by adding four kinds of organic solvents at the concentration level of 4%(volume fraction

  17. EFEITO DO TEOR DE GORDURA, VÁCUO E DOSE DE RADIAÇÃO GAMA NA SOBREVIVÊNCIA DE Salmonella TYPHIMURIUM ATCC 14028 EM CARNE BOVINA MOÍDA RESFRIADA

    Directory of Open Access Journals (Sweden)

    C. S. COSTA

    2008-09-01

    Full Text Available

    O trabalho avaliou a sobrevivência de Salmonella Typhimurium ATCC 14028 em carnes bovinas, moída crua e resfriada (2 ºC, através do tratamento com radiação gama (Co60, utilizando doses de 0; 1,5; 2,5 e 3,5 kGy. Além do fator dose de radiação foram avaliadas as influências do emprego de vácuo e de dois teores de gordura da carne bovina moída: baixo (2-4% e alto (11-13%, bem como a interação dos fatores, na redução ou eliminação da bactéria patogênica inoculada. Os resultados demonstraram que os teores de gordura da carne e o emprego de vácuo não influenciaram significativamente a sobrevivência da Salmonella. A dose de radiação gama influenciou a inativação de Salmonella de forma dose dependente até 2,5 kGy, com reduções de 4 ciclos logarítmicos. A dose de 2,5 kGy é suficiente para exercer um controle efetivo de Salmonella em carne bovina moída independentemente do seu teor de gordura e da presença de oxigênio.

  18. The Effect of Lactobacillus plantarum ATCC 8014 and Lactobacillus acidophilus NCFM Fermentation on Antioxidant Properties of Selected in Vitro Sprout Culture of Orthosiphon aristatus (Java Tea as a Model Study

    Directory of Open Access Journals (Sweden)

    Dase Hunaefi

    2012-09-01

    Full Text Available High rosmarinic acid (RA productivity has been achieved by applying jasmonic acid and yeast extract elicitors to the in vitro sprout culture of Orthosiphon aritatus (IOSC. The highest RA accumulation from three solvents was detected in IOSC after treatment with yeast extract (5 g/L. HPLC analysis clearly confirmed a drastic increase in RA subjected to yeast extract elicitation. Therefore, this yeast extract elicited IOSC was chosen for a lactic acid bacteria (LAB fermentation study as a model system. This selected IOSC was subjected to different types of LAB fermentations (Lactobacillus plantarum ATCC 8014 and Lactobacillus acidophilus NCFM for different periods of time 24, 48 and 72 h. The LAB fermentations consisted of solid state fermentations (SSF and liquid state fermentations (LSF in a Digital Control Unit (DCU fermenter system. The aim was to determine the effect of fermentation on the antioxidant properties of the plant extract. Results indicated that all types of LAB fermentation decreased the level of RA and total phenolics, however, a slight increase in total flavonoids and flavonols was observed in SSF samples. HPLC results confirmed that the longer the fermentation, the greater the reduction in RA content. The highest reduction was obtained in the sample of LSF inoculated with L. plantarum for a period of 72 h. The temperature of fermentation (37 °C was predicted as contributing to the declining level in RA content. The loss in RA was concomitant with a loss of total antioxidant activity (1,1-diphenyl-2-picrylhydrazyl (DPPH scavenging activity, Trolox Equivalent Antioxidant Capacity (TEAC, and Superoxide Dismutase (SOD-like activity. These results indicate that RA is the major contributor to the antioxidant activity of this plant.

  19. The Effect of pH Control on Acetone-Butanol-Ethanol Fermentation by Clostridium acetobutylicum ATCC 824 with Xylose and D-Glucose and D-Xylose Mixture

    Institute of Scientific and Technical Information of China (English)

    Wei Jiang; Zhiqiang Wen; Mianbin Wu; Hong Li; Jun Yang; Jianping Lin; Yijun Lin; Lirong Yang; Peilin Cen

    2014-01-01

    D-Glucose, L-arabinose, D-mannose, D-xylose, and cellobiose are saccharification products of lignocellulose and important carbon sources for industrial fermentation. The fermentation efficiency with each of the five sugars and the mixture of the two most dominant sugars, D-glucose and D-xylose, was evaluated for acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum ATCC 824. The utilization efficacy of the five reducing sugars was in the order of D-glucose, L-arabinose, D-mannose, D-xylose and cellobiose. D-Xylose, the second most abundant component in lignocellulosic hydrolysate, was used in the fermentation either as sole carbon source or mixed with glucose. The results indicated that maintaining pH at 4.8, the optimal pH value for solventogenesis, could increase D-xylose consumption when it was the sole carbon source. Different media con-taining D-glucose and D-xylose at different ratios (1:2, 1:5, 1.5:1, 2:1) were then attempted for the ABE fermenta-tion. When pH was at 4.8 and xylose concentration was five times that of glucose, a 256.9%increase in xylose utilization and 263.7%increase in solvent production were obtained compared to those without pH control. These results demonstrate a possible approach combining optimized pH control and D-glucose and D-xylose ratio to increase the fermentation efficiency of lignocellulosic hydrolysate.

  20. Role of transcription and enzyme activities in redistribution of carbon and electron flux in response to N₂ and H₂ sparging of open-batch cultures of Clostridium thermocellum ATCC 27405.

    Science.gov (United States)

    Carere, Carlo R; Rydzak, Thomas; Cicek, Nazim; Levin, David B; Sparling, Richard

    2014-03-01

    Growth, end-product synthesis, enzyme activities, and transcription of select genes associated with the "malate shunt," pyruvate catabolism, H2 synthesis, and ethanol production were studied in the cellulolytic anaerobe, Clostridium thermocellum ATCC 27405, during open-batch fermentation of cellobiose to determine the effect of elevated N2 and H2 gas sparging on metabolism using a 14-L fermenter with a 7-L working volume. The metabolic shift from acetate, H2, and CO2 to ethanol and formate in response to high H2 versus high N2 sparging (20 mL s(-1)) was accompanied by (a) a 2-fold increase in nicotinamide adenine dinucleotide (NADH)-dependent alcohol dehydrogenase (Adh) activity, (b) a 10-fold increase in adhE transcription, and (c) a 3-fold decrease in adhZ transcription. A similar, but less pronounced, metabolic shift was also observed when the rate of N2 sparging was decreased from 20 to 2 mL s(-1), during which (a) NADH-dependent ADH and pyruvate: ferredoxin oxidoreductase (PFOR) activities increased by ∼1.5-fold, (b) adhY transcription increased 6-fold, and (c) transcription of selected pfor genes increased 2-fold. Here we demonstrate that transcription of genes involved in ethanol metabolism is tightly regulated in response to gas sparging. We discuss the potential impacts of dissolved H2 on electron carrier (NADH, NADPH, ferredoxin) oxidation and how these electron carriers can redirect carbon and electron flux and regulate adhE transcription. PMID:24463715

  1. Molecular analysis and anticancer properties of two identified isolates, Fusarium solani and Emericella nidulans isolated from Wady El-Natron soil in Egypt against Caco-2 (ATCC) cell line

    Institute of Scientific and Technical Information of China (English)

    Hala F Mohamed

    2012-01-01

    Objective: To characterize, identify and investigate the anticancer properties of two new soil fungal isolates, Emericella nidulans and Fusarium solani isolated from Wady El-Natron in Egypt against colon cancer Caco-2 (ATCC) cell line. Methods: Soil sample was cultured and two strains were chosen for morphological and phenotypical characterization. Partial sequences of the 18s rRNA gene and the internal transcribed spacer region ITS of the two isolates were amplified by PCR. Phylogenetic tree construction and analysis of the resulted multiple sequences from the two fugal isolates were also carried out. In vitro anticancer activity of the two strains was done against colon Caco-2 cancer cell line. Reverse transcription – PCR was carried out to detect level of expression of p53 in Caco-2 cell line. Results: HF.1 displayed morphological and genotypic characteristics most similar to that of Fusarium solani while HF.2 was most similar to Emericella nidulans with high similarity of 99% and 97% respectively. The multiple sequence alignment of the two fungal isolates showed that, the maximum identical conserved domains in the 18s rRNA genes were identified with the nucleotide regions of 51st to 399th base pairs, 88th to 525th base pairs respectively. While those in the ITS genes were identified with the nucleotide regions of 88th to 463rdand 51st to 274th. The two isolates showed IC50 value with (6.24±5.21) and (9.84±0.36) μg/mL) concentrations respectively at 28h. Reverse transcription – PCR indicated that these cells showed high level of expression for p53 mRNA. Conclusions: The morphology and molecular analysis identified HF.1 and HF.2 to be Fusarium solani and Emericella nidulans; new isolates of anticancer producing fungi from Wady El-Natroon city in Egypt. Treatment with the two isolates caused P53 expression in Caco-2 cell line. These two isolates can be used as an anticancer agents.

  2. Biosíntesis de dextranos de alto peso molecular mediante la inoculación con Leuconostoc Mesenteroides, var. Mesenteroides (atcc 10830 de jugos residuales de la agroindustria de la piña: síntesis y caracterización de hierro-dextranos

    Directory of Open Access Journals (Sweden)

    José Roberto Vega Baudrit

    2013-01-01

    Full Text Available En este trabajo se muestran los estudios realizados para obtener dextranos a partir de desechos de la agroindustria de piña. La fermentación se llevó a cabo en un biorreactor (10 L, se inoculó con un cultivo de Leuconostoc mesenteroides, var. mesenteroides (ATCC 10830. Se centrifugó y se precipitó y purificó con etanol. Fue caracterizado por medio de viscosidad, peso molecular y grupos funcionales por espectroscopía infrarroja. Este dextrano fue tratado con el fin de obtener hierro-dextranos.

  3. Application of thermoalkalophilic xylanase from Arthrobacter sp. MTCC 5214 in biobleaching of kraft pulp

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Bhosle, N.B.

    released by enzyme treatment showed a characteristic peak at 280 nm indicating the presence of lignin in the released coloring matter. Enzymatic prebleaching of kraft pulp showed 20 % reduction in kappa number of the pulp without much change in viscosity...

  4. CLONING AND PURIFICATION OFA REPRESSOR PROTEIN FROM ARTHROBACTER NICOTINOVORANS PAO1

    Directory of Open Access Journals (Sweden)

    Marius Mihasan

    2010-09-01

    Full Text Available The pAO1 megaplasmid of A. nicotinovorans consists of 165 ORF's related mainly to nicotine degradation, uptake and utilization of carbohydrates, amino acids and sarcosine The putative sugar catabolic pathway consists of 11 ORFś organized as a single operon and coding for an ABC-type sugar-transport system and several putative oxidoreductases and dehydrogenases. The current work is focused on orf32, a putative PdhR related protein, most probably involved in the control of the whole operon. The approx. 700 kb orf32 gene was cloned in the pH6EX3 plasmid vector and the gene product purified to homogeneity as a 29 kDa His-tagged recombinant protein. As indicated by GPC, it consists of a monomeric protein with a native molecular weight of 32 kDa. The specific UV/Vis spectra showed only a single peak at 280 nm common for all proteins and did not indicated the presence of any colored cofactors. This is in good agreement with the fact that PdhR-family proteins contain a winged helix-turn-helix (wHTH domain responsible for DNA binding, and not a Zn-finger or any other metal containing domains

  5. Purification and characterization of a novel marine Arthrobacter oxydans KQ11 dextranase.

    Science.gov (United States)

    Wang, Delong; Lu, Mingsheng; Wang, Shujun; Jiao, Yuliang; Li, Weijuan; Zhu, Qiang; Liu, Zhaopu

    2014-06-15

    Dextranases can hydrolyze dextran deposits and have been used in the sugar industry. Microbial strains which produce dextranases for industrial use are chiefly molds, which present safety issues, and dextranase production from them is impractically long. Thus, marine bacteria to produce dextranases may overcome these problems. Crude dextranase was purified by a combination of ammonium sulfate fractionation and ion-exchange chromatography, and then the enzyme was characterized. The enzyme was 66.2 kDa with an optimal temperature of 50°C and a pH of 7. The enzyme had greater than 60% activity at 60°C for 1h. Moreover, 10mM Co(2+) enhanced dextranase activity (196%), whereas Ni(2+) and Fe(3+) negatively affected activity. 0.02% xylitol and 1% alcohol enhanced activity (132.25% and 110.37%, respectively) whereas 0.05% SDS inhibited activity (14.07%). The thickness of S. mutans and mixed-species oral biofilm decreased from 54,340 nm to 36,670 nm and from 64,260 nm to 43,320 nm, respectively. PMID:24721052

  6. Effect of Arthrobacter agilis UMCV2 on germination and growth of Pinus devoniana Lindley

    Directory of Open Access Journals (Sweden)

    Wilber Montejo-Mayo

    2016-03-01

    Full Text Available Soil microorganisms are essential for growth, emergence and development in all plants. In our study we decided to evaluate the effect that A. agilis UMCV2 rhizobacteria had on germination and growth of plants of economic-forest interest as P. devoniana to an age of 65 days. Our results show that the UMCV2 bacteria promoted growth of P. devoniana at this early stage of development, further highlighting that despite the short time of interaction between these two, the bacterium was able to increase the rate of germination, increase the size of shoot and generate a proliferation of lateral roots. The data shows a huge potential for using inoculum both in the greenhouse and in open ground and generate a growth promoting species of interest in both agricultural and forest, thereby reducing production periods depending on the crop.

  7. Cloning of genes and developing transgenic crops with enhanced tolerance to salinity and drought (abstract)

    International Nuclear Information System (INIS)

    Abiotic stresses represent the most limiting factors affecting agricultural productivity. In India more than 60% of total cultivated land is still rainfed and crops experience frequent droughts. Thus, we need to develop transgenic crops tolerant to drought, and other related abiotic stress factors such as salinity, low and high temperature stresses. At the National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute (ICAR), we have initiated a programme on developing transgenic crops tolerant to a range of abiotic stresses. The major emphasis is on developing transgenic potato, tomato, mustard, rice and wheat. While, transgenic plants of potato. tomato and mustard have already been generated with osmotin gene and are at different stages of testing, other key genes imparting tolerance to abiotic stresses are being isolated from different species for producing transgenic rice and wheat cultivars tolerant to multiple stresses. Genes that have been isolated in our laboratory include ascorbate peroxidase gene (TaApx) and genes encoding transcription factor, CBFs (TaCBF2 and TaCBP3) from a drought tolerant wheat cultivar (C306), Lea1 cDNA from Brassica species, codA from Arthrobacter globiformis, and otsBA operon from E. coli. Apart from these stress-related genes, we have isolated a few stress-inducible promoters for deploying them in gene stacking in developing transgenic crops with enhanced tolerance to multiple abiotic stresses. The results will be presented. (author)

  8. Variability of sediment-contact tests in freshwater sediments with low-level anthropogenic contamination - Determination of toxicity thresholds

    Energy Technology Data Exchange (ETDEWEB)

    Hoess, S., E-mail: hoess@ecossa.d [Ecossa, Giselastr. 6, 82319 Starnberg (Germany); Institute of Biodiversity - Network (IBN), Dreikronengasse 2, 93047 Regensburg (Germany); Ahlf, W., E-mail: ahlf@tu-harburg.d [Institute of Environmental Technology and Energy Economics, Technical University Hamburg-Harburg, Eissendorfer Str. 40, 21071 Hamburg (Germany); Fahnenstich, C. [Institute of Environmental Technology and Energy Economics, Technical University Hamburg-Harburg, Eissendorfer Str. 40, 21071 Hamburg (Germany); Gilberg, D., E-mail: d-gilberg@ect.d [ECT Oekotoxikologie, Boettgerstr. 2-14, 65439 Floersheim (Germany); Hollert, H., E-mail: henner.hollert@bio5.rwth-aachen.d [Department of Ecosystem Analysis, Institute for Environmental Research (Biology 5), RWTH Aachen University, Worringerweg 1, 52074 Aachen (Germany); Melbye, K. [Dr. Fintelmann and Dr. Meyer, Mendelssohnstr. 15D, 22761 Hamburg (Germany); Meller, M., E-mail: m-meller@ecotox-consult.d [ECT Oekotoxikologie, Boettgerstr. 2-14, 65439 Floersheim (Germany); Hammers-Wirtz, M., E-mail: hammers-wirtz@gaiac.rwth-aachen.d [Research Institute for Ecosystem Analysis and Assessment (gaiac), RWTH Aachen University, Worringerweg 1, 52056 Aachen (Germany); Heininger, P., E-mail: heininger@bafg.d [Federal Institute of Hydrology (BfG), Am Mainzer Tor 1, 56070 Koblenz (Germany); Neumann-Hensel, H., E-mail: hensel@fintelmann-meyer.d [Dr. Fintelmann and Dr. Meyer, Mendelssohnstr. 15D, 22761 Hamburg (Germany); Ottermanns, R., E-mail: ottermanns@bio5.rwth-aachen.d [Chair for Environmental Biology and Chemodynamics, Institute for Environmental Research (Biology 5), RWTH Aachen University, Worringerweg 1, 52074 Aachen (Germany); Ratte, H.-T., E-mail: toni.ratte@bio5.rwth-aachen.d [Chair for Environmental Biology and Chemodynamics, Institute for Environmental Research (Biology 5), RWTH Aachen University, Worringerweg 1, 52074 Aachen (Germany)

    2010-09-15

    Freshwater sediments with low levels of anthropogenic contamination and a broad range of geochemical properties were investigated using various sediment-contact tests in order to study the natural variability and to define toxicity thresholds for the various toxicity endpoints. Tests were performed with bacteria (Arthrobacter globiformis), yeast (Saccharomyces cerevisiae), nematodes (Caenorhabditis elegans), oligochaetes (Lumbriculus variegatus), higher plants (Myriophyllum aquaticum), and the eggs of zebrafish (Danio rerio). The variability in the response of some of the contact tests could be explained by particle size distribution and organic content. Only for two native sediments could a pollution effect not be excluded. Based on the minimal detectable difference (MDD) and the maximal tolerable inhibition (MTI), toxicity thresholds (% inhibition compared to the control) were derived for each toxicity parameter: >20% for plant growth and fish-egg survival, >25% for nematode growth and oligochaete reproduction, >50% for nematode reproduction and >60% for bacterial enzyme activity. - Sediment-contact tests require toxicity thresholds based on their variability in native sediments with low-level contamination.

  9. Variability of sediment-contact tests in freshwater sediments with low-level anthropogenic contamination - Determination of toxicity thresholds

    International Nuclear Information System (INIS)

    Freshwater sediments with low levels of anthropogenic contamination and a broad range of geochemical properties were investigated using various sediment-contact tests in order to study the natural variability and to define toxicity thresholds for the various toxicity endpoints. Tests were performed with bacteria (Arthrobacter globiformis), yeast (Saccharomyces cerevisiae), nematodes (Caenorhabditis elegans), oligochaetes (Lumbriculus variegatus), higher plants (Myriophyllum aquaticum), and the eggs of zebrafish (Danio rerio). The variability in the response of some of the contact tests could be explained by particle size distribution and organic content. Only for two native sediments could a pollution effect not be excluded. Based on the minimal detectable difference (MDD) and the maximal tolerable inhibition (MTI), toxicity thresholds (% inhibition compared to the control) were derived for each toxicity parameter: >20% for plant growth and fish-egg survival, >25% for nematode growth and oligochaete reproduction, >50% for nematode reproduction and >60% for bacterial enzyme activity. - Sediment-contact tests require toxicity thresholds based on their variability in native sediments with low-level contamination.

  10. Appraising bacterial strains for rapid BOD sensing--an empirical test to identify bacterial strains capable of reliably predicting real effluent BODs.

    Science.gov (United States)

    Webber, Judith B; Noonan, Mike; Pasco, Neil F; Hay, Joanne M

    2011-01-01

    The measured response of rapid biochemical oxygen demand (BOD) biosensors is often not identical to those measured using the conventional 5-day BOD assay. This paper highlights the efficacy of using both glucose-glutamic acid (GGA) and Organisation for Economic Cooperation and Development (OECD) BOD standards as a rapid screen for microorganisms most likely to reliably predict real effluent BODs when used in rapid BOD devices. Using these two synthetic BOD standards, a microorganism was identified that produced comparable BOD response profiles for two assays, the MICREDOX® assay and the conventional 5-day BOD(5) test. A factorial experimental design systematically evaluated the impact of four factors (microbial strain, growth media composition, media strength, and microbial growth phase) on the BOD response profiles using GGA and OECD synthetic standard substrates. An outlier was identified that showed an improved correlation between the MICREDOX® BOD (BOD(sens)) and BOD(5) assays for both the synthetic standards and for real wastewater samples. Microbial strain was the dominant factor influencing BOD(sens) values, with Arthrobacter globiformis single cultures clearly demonstrating superior rapid BOD(sens) response profiles for both synthetic and real waste samples. It was the only microorganism to approach the BOD(5) response for the OECD substrate (171 mg O(2)L(-1)), and also reported BOD values for real waste samples that were comparable to those produced by the BOD(5) test, including discriminating between filtered and unfiltered samples.

  11. Production of interleukin-1β, interleukin-6 and tumor necrosis factor α in cultured human fibroblast with stimulation of abstract from Actinomyces naeslundii ATCC19246%内氏放线菌细胞壁成分诱导人成纤维细胞产生白细胞介素-1β、-6和肿瘤坏死因子α的研究

    Institute of Scientific and Technical Information of China (English)

    赵丽娟; 李文; 郑燕

    2008-01-01

    目的 探讨内氏放线菌菌株ATCC19246细胞壁成分能否诱导人成纤维细胞产生白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子α(TNFα).方法 采用提取脂磷壁酸(LTA)的方法 提取内氏放线菌菌株ATCC19246细胞壁成分,用3种质量浓度(分别为1、10、100 mg/mL)提取物分别刺激人成纤维细胞THP-1细胞株,用ELISA方法 测定细胞培养上清中的IL-1β、IL-6、TNFα.结果 THP-1细胞株可以产生一定量的IL-1β、IL-6、TNFα,以质量浓度为10 mg/mL时诱导产生的量最高.结论 内氏放线菌细胞壁提取物可以诱导THP-1细胞株产生IL-1β、IL-6、TNFα,但随着质量浓度变化产生的量存在差异.

  12. Metabolism of methoxychlor by Cunninghamella elegans ATCC36112.

    Science.gov (United States)

    Keum, Young Soo; Lee, Youn Hyung; Kim, Jeong-Han

    2009-09-01

    Methoxychlor is considered as pro-estrogen, while some of its metabolites are more potent endocrine disruptors than the parent insecticide. Major activation of methoxychlor is through cytochrome P450-catalyzed demethylation to bisphenol A-like metabolites. Cunninghamella elegans is a well-known fungal species with its strong resemblance of the xenobiotic metabolism of the mammalian system. In this study, the metabolism of methoxychlor was investigated with the corresponding organism. Methoxychlor was rapidly transformed to approximately 11 metabolites in phase I metabolism, including oxidation, hydroxylation, and dechlorination. Concentrations of phase I metabolites reached a maximum at 4-6 days and gradually decreased until the end of the experiments. Most metabolites from the phase I reaction were further transformed to sugar conjugates. Approximately 11 or more glucose conjugates were found in culture supernatants and gradually increased, while no glucuronides were observed throughout the experiments. Piperonyl butoxide and chlorpyrifos strongly inhibit the degradation of methoxychlor and concomitant accumulation of metabolites, indicating cytochrome P450 mediated metabolism. Little or no glycosides were detected in chlorpyrifos- and piperonyl butoxide-treated cultures. From the results, Cunninghamella elegans has shown strong similarities of the phase I metabolism of methoxychlor, while the conjugation reaction is different from those of animal metabolism.

  13. Genome Sequence of the Pathogenic Fungus Sporothrix schenckii (ATCC 58251).

    Science.gov (United States)

    Cuomo, Christina A; Rodriguez-Del Valle, Nuri; Perez-Sanchez, Lizaida; Abouelleil, Amr; Goldberg, Jonathan; Young, Sarah; Zeng, Qiandong; Birren, Bruce W

    2014-05-22

    Sporothrix schenckii is a pathogenic dimorphic fungus that grows as a yeast and as mycelia. This species is the causative agent of sporotrichosis, typically a skin infection. We report the genome sequence of S. schenckii, which will facilitate the study of this fungus and of the Sporothrix schenckii group.

  14. Antibacterial Characterization of Silver Nanoparticles against E. Coli ATCC-15224

    Institute of Scientific and Technical Information of China (English)

    M.Raffi; F.Hussain; T.M.Bhatti; J.I.Akhter; A.Hameed; M.M.Hasan

    2008-01-01

    Silver nanoparticles of mean size 16 nm were synthesized by inert gas condensation (IGC) method. Crystalline structure, morphology and nanoparticles size estimation were conducted by X-ray diffraction (XRD) and transmission electron microscopy (TEM). Antibacterial activity of these silver nanoparticles as a function of particles concentration against gram-negative bacterium Escherichia coli (E. coli) was carried out in liquid as well as solid growth media. Scanning electron microscopy (SEM) and TEM studies showed that silver nanoparticles after interaction with E.coli have adhered to and penetrated into the bacterial cells. Antibacterial properties of silver nanoparticles are attributed to their total surface area, as a larger surface to volume ratio of nanoparticles provides more efficient means for enhanced antibacterial activity.

  15. Induction of natural competence in Bacillus cereus ATCC14579

    NARCIS (Netherlands)

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

    2008-01-01

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

  16. Colonization of Crystalline Cellulose by Clostridium cellulolyticum ATCC 35319

    OpenAIRE

    Gelhaye, E.; Gehin, A; Petitdemange, H.

    1993-01-01

    Cellulose colonization by Clostridium cellulolyticum was studied by using [methyl-3H]thymidine incorporation. The colonization process indicated that a part of the bacterial population was released from cellulose to the liquid phase before binding and colonizing another adhesion site of the cellulose. We postulate that cellulose colonization occurs according to the following process: adhesion, colonization, release, and readhesion.

  17. 睾丸酮丛毛单胞菌ATCC 11996类固醇脱氢酶3α-HSD的高效表达与纯化%Effective expression and purification of 3α-HSD protein from Comamonas testosteroni

    Institute of Scientific and Technical Information of China (English)

    肖琳; 薛强; 邹明强; 冯叙桥; 韦娜; 何田田; 孙芳芳

    2013-01-01

    3α-HSD gene was amplified and cloned into the expression vector pET-15b. The constructed plasmid was over expressed in the host bacterial of E. Coli BL21 after IPTG induction. Expression of 3α-HSD was optimized by bacterial culture temperature,concentration of IPTG and induction time. The engineering bacteria was lysed,and purified by ammounium sulfate precipitation, Ni-NTA affinity purification and molecular sieve separation. Enzymatic activity of expressed 3a-HSD was detected. Comamonas testosteroni 3a-HSD gene was cloned and expressed in the prokaryotic system. To reach the optimal expression, the recombinant E. Coli need to be cultured at 37 ℃ and induced by 0. 6 mmol/L IPTG for 4 h, while the medium containing 0.1 mmol/L Zn2+ . The protein was purified through 60% ammonium sulfate precipitation, Ni- NTA affinity chromatography, and molecular sieve. A target protein with 99% purity was obtained which keeping enzyme activity.3α-HSD gene was cloned and highly expressed. It will build the foundation of antibody preparation and steroid detection in the water resource.%利用分子克隆的方法扩增与克隆睾丸酮丛毛单胞菌ATCC 11996的3α-HSD基因,构建以pET-15b为载体的表达工程菌,IPTG诱导蛋白表达,通过优化表达菌的最佳表达条件得到最大表达量的目的蛋白.诱导后将工程菌裂解,通过硫酸铵沉淀,Ni-NTA亲和层析,分子筛分离进行纯化,并测定纯化后蛋白的酶活性.结果表明:1)克隆获得了睾丸酮丛毛单胞菌3α-HSD基因并实现了异源表达;2)工程菌的最佳表达条件为:37℃培养,0.6mmol/L IPTG诱导4h,培养基中Zn2+终浓度为0.1 mmol/L;3)表达工程菌裂解后用60%硫酸铵沉淀,亲和层析和分子筛分离后得到了纯度高达99%的目的蛋白,纯化后的蛋白保持酶活性.本试验成功克隆表达了睾丸酮丛毛单胞菌3α-HSD基因,建立了3α-HSD的高效表达与纯化方法,为相应抗体的制备与水资源中的类固醇激

  18. Class 1 integrons and tetracycline resistance genes in Alcaligenes, Arthrobacter, and Pseudomonas spp. isolated from pigsties and manured soil

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Sandvang, Dorthe

    2005-01-01

    The presence of tetracycline resistance (Tc-r) genes and class I integrons (in-1), and their ability to cotransfer were investigated in Tc-r gram-negative (185 strains) and gram-positive (72 strains) bacteria from Danish farmland and pigsties. The isolates belonged to the groups or species...... tet(33). No isolates contained more than one tet gene. The in-l-positive isolates were tested for resistance to selected antimicrobial agents and showed resistance to three to nine drugs. Filter-mating experiments showed cotransfer of Tc-r and class I integrons from soil isolates to Escherichia coli...... and/or Pseudomonas putida. We conclude that soil bacteria in close contact to manure or pigsty environment may thus have an important role in horizontal spread of resistance. Use of tetracyclines in food animal production may increase not only Tc-r but also multidrug resistance (caused by the presence...

  19. Characterizing the Catalytic Potential of Deinococcus, Arthrobacter and other Robust Bacteria in Contaminated Subsurface Environments of the Hanford Site

    Energy Technology Data Exchange (ETDEWEB)

    Daly, Michael J.

    2007-07-23

    Progress is briefly summarized in these areas: ionizing radiation resistance in bacteria; a hypothesis regarding ionizing radiation resistance emerging for bacterial cells; transcriptome analysis of irradiated D. radiodurans and Shewanella oneidensis; the role of metal reduction in Mn-dependnet Deinococcal species; and engineered Deinococcus strains as models for bioremediation. Key findings are also reported regarding protein oxidation as a possible key to bacterial desiccation resistance, and the whole-genome sequence of the thermophile Deinococcus geothermalis.

  20. MOLECULAR GENE CLONING OF NICOTINE-DEHIDROGENASE FROM THE pAO1 MEGAPLASMID OF ARTHROBACTER NICOTINOVORANS

    Directory of Open Access Journals (Sweden)

    Andreea Andrei

    2013-10-01

    Full Text Available 6-hydroxi-L-nicotine (6HNic has an important potential as a drug for neuro-degenerative disorders and a  suitable simple and reliable method for obtaining contaminant-free 6HNic preparations is required. Here, we envision the in-vitro production of 6HNic by using purified nicotine-dehydrogenase (NDH followed by HPLC or capillary electrophoresis techniques and we focus on the isolation and cloning of the three genes coding the NDH enzyme.  A PCR protocol was established for easy amplification and the DNA fragment containing the ndhLSM genes was directionally cloned into the pART2 vector.

  1. Experimental evidence of a xylose-catabolic pathway on the pAO1 megaplasmid of Arthrobacter nicotinovorans

    Directory of Open Access Journals (Sweden)

    Marius Mihasan

    2012-09-01

    Full Text Available The pAO1 megaplasmid of A. nicotinovorans consists of 165 ORF's related mainly to nicotine degradation, uptake and utilization of carbohydrates, amino acids and sarcosine. A putative sugar catabolic pathway consisting of 11 ORF's organized as a single operon were previously described. The current work brings experimental data supporting the existence of a D-Xylose catabolic pathway on the pAO1 megaplasmid. When grown on D-xylose containing media, the cells harboring the pAO1 megaplasmid grow to higher cell densities and also express the OxRe protein coded by the megaplasmid. A putative pathway similar to Weimberg pentose pathway is postulated, in which D-xylose is transported in the cell by the ABC-type transport system and then transformed using the putative sugar-dehidrogenase OxRe to D-xylonate, which is further degrated to 2-ketoglutarate and integrated into the general metabolism of the cell

  2. Swapping metals in Fe- and Mn-dependent dioxygenases: Evidence for oxygen activation without a change in metal redox state

    Energy Technology Data Exchange (ETDEWEB)

    Emerson, Joseph P.; Kovaleva, Elena G.; Farquhar, Erik R.; Lipscomb, John D.; Oue, Jr., Lawrence (UMM)

    2008-07-21

    Biological O{sub 2} activation often occurs after binding to a reduced metal [e.g., M(II)] in an enzyme active site. Subsequent M(II)-to-O{sub 2} electron transfer results in a reactive M(III)-superoxo species. For the extradiol aromatic ring-cleaving dioxygenases, we have proposed a different model where an electron is transferred from substrate to O{sub 2} via the M(II) center to which they are both bound, thereby obviating the need for an integral change in metal redox state. This model is tested by using homoprotocatechuate 2,3-dioxygenases from Brevibacterium fuscum (Fe-HPCD) and Arthrobacter globiformis (Mn-MndD) that share high sequence identity and very similar structures. Despite these similarities, Fe-HPCD binds Fe(II) whereas Mn-MndD incorporates Mn(II). Methods are described to incorporate the nonphysiological metal into each enzyme (Mn-HPCD and Fe-MndD). The x-ray crystal structure of Mn-HPCD at 1.7 {angstrom} is found to be indistinguishable from that of Fe-HPCD, while EPR studies show that the Mn(II) sites of Mn-MndD and Mn-HPCD, and the Fe(II) sites of the NO complexes of Fe-HPCD and Fe-MndD, are very similar. The uniform metal site structures of these enzymes suggest that extradiol dioxygenases cannot differentially compensate for the 0.7-V gap in the redox potentials of free iron and manganese. Nonetheless, all four enzymes exhibit nearly the same K{sub M} and V{sub max} values. These enzymes constitute an unusual pair of metallo-oxygenases that remain fully active after a metal swap, implicating a different way by which metals are used to promote oxygen activation without an integral change in metal redox state.

  3. Potential application of metabolic engineering to tune the production of compatible solutes for enhancing tolerance of crop plants to salinity/drought (abstract)

    International Nuclear Information System (INIS)

    Essential need to develop genotypes of crop plants that can substantially withstand salinity and drought with little yield losses is being increasingly felt, as the cultivable agricultural lands is increasingly being exposed to these stresses. In-spite of gains in productivity, conventional plant breeding methods have their limitations either due to limited gene pool or due to species barrier for gene transfer. Modern molecular tools have paved ways for identification of genes imparting abiotic stress tolerance in unrelated species/organisms and to transfer the selected genes into desirable crop plant species by conquering the incompatibility barriers. In fact, now genetic engineering has been widely realized to be in important tool for developing abiotic stress tolerant crop plants. Abiotic stress tolerance is a complex phenomenon involving simultaneous expression of a number of genes coupled with an interaction of varying weather variables and crop phonology. However, in order to tackle the issue, successful attempts have been made in identifying genes enhancing abiotic stress tolerance. The genes for biosynthesis of various compatible solutes (viz., mtlD for mannitol: P5CS or P5CSF129A for proline; coda/cox or belA/beIB for glycinebetaine' lpsl for trehalose; PINOI for inositol) have been demonstrated to enhance abiotic stress tolerance of plants. We have isolated the codA gene (Accession number AY589052) for choline oxidase from an Indian strain of Arthrobacter sp. from IMTECH (Chandigarh) and the mtlD genes from local strains of E. coli (accession number A Y523630) and halobacterium sp. (Accession number A Y52363 1). We have enhanced the tolerance of Brassica juncea to salt, drought and low temperature stresses by introducing the codA gene from Arthrobacter globiformis using Agrobacterium tumefaciens mediated transformation. Presenting our research team is busy developing genotypes of chickpea black gram, peanut and sorghum besides mustard with enhanced

  4. Rizoremediação de pentaclorofenol em um solo argiloso por sphingomonas chlorophenolica ATCC 39723 Pentachlorophenol rhizoremediation in a loamy soil by sphingomonas chlorophenolica ATCC 39723

    Directory of Open Access Journals (Sweden)

    Rosemeri I. Dams

    2007-12-01

    Full Text Available O principal objetivo deste trabalho foi estudar a degradação de PCP por Sphingomonas chlorophenolicaem solo argiloso na presença e ausência de trigo. As concentrações de PCP foram determinadas através de Análises de Alta Performance de Cromatografia Líquida. Os efeitos tóxicos de PCP foram estudados através do monitoramento do crescimento das plantas. A biodegradação de PCP por S. chlorophenolica foi acompanhada por testes de bioluminescência de Escherichia coli HB101 pUCD607 e contagens bacterianas no solo e nas raízes. A degradação de PCP ocorreu de forma mais rápida no solo plantado e inoculado quando comparada ao solo sem plantas. Houve um aumento significativo nas populações dos organismos testados nas raízes quando comparadas com as populações presentes no solo. O monitoramento do crescimento da planta mostrou o papel protetor exercido pela S.chlorophenolica contra a toxicidade do PCP.The main objective of this study was study the PCP degradation by Sphingomonas chlorophenolica in a loamy soil in the presence and absence of plants (Winter wheat. Measurements of PCP concentrations were carried out in a laboratory basis using High performance liquid chromatography analysis (HPLC. The toxic effect of PCP on plants was studied through the monitoring of the plant growth. The biodegradation of PCP by S. chlorophenolica in soil was assessed with a bioluminescence assay of Escherichia coli HB101 pUCD607 and bacterial analyses in roots and soil. The planted and inoculated soil showed a faster degradation when compared to the inoculated soil without plants. There was a significative increase in the populations of the organisms tested in the roots when compared to the soil. The monitoring of the plant growth showed a protective role of S. chlorophenolica against the toxicity of PCP in the loamy soil.

  5. Fusobacterium nucleatum ATCC 10953 requires Actinomyces naeslundii ATCC 43146 for growth on saliva in a three-species community that includes Streptococcus oralis 34.

    Science.gov (United States)

    Periasamy, Saravanan; Chalmers, Natalia I; Du-Thumm, Laurence; Kolenbrander, Paul E

    2009-05-01

    Formation of dental plaque is a developmental process involving initial and late colonizing species that form polymicrobial communities. Fusobacteria are the most numerous gram-negative bacteria in dental plaque, but they become prevalent after the initial commensal colonizers, such as streptococci and actinomyces, have established communities. The unusual ability of these bacteria to coaggregate with commensals, as well as pathogenic late colonizers, has been proposed to facilitate colonization by the latter organisms. We investigated the integration of Fusobacterium nucleatum into multispecies communities by employing two in vitro models with saliva as the sole nutritional source. In flow cell biofilms, numbers of cells were quantified using fluorescently conjugated antibodies against each species, and static biofilms were analyzed by quantitative real-time PCR (q-PCR) using species-specific primers. Unable to grow as single-species biofilms, F. nucleatum grew in two-species biofilms with Actinomyces naeslundii but not with Streptococcus oralis. However, enhanced growth of fusobacteria was observed in three-species biofilms, indicating that there was multispecies cooperation. Importantly, these community dynamics yielded an 18-fold increase in the F. nucleatum biomass between 4 h and 18 h in the flow cell inoculated with three species. q-PCR analysis of static biofilms revealed that maximum growth of the three species occurred at 24 h to 36 h. Lower numbers of cells were observed at 48 h, suggesting that saliva could not support higher cell densities as the sole nutrient. Integration of F. nucleatum into multispecies commensal communities was evident from the interdigitation of fusobacteria in coaggregates with A. naeslundii and S. oralis and from the improved growth of fusobacteria, which was dependent on the presence of A. naeslundii. PMID:19286780

  6. Fusobacterium nucleatum ATCC 10953 Requires Actinomyces naeslundii ATCC 43146 for Growth on Saliva in a Three-Species Community That Includes Streptococcus oralis 34▿

    OpenAIRE

    Periasamy, Saravanan; Chalmers, Natalia I.; Du-Thumm, Laurence; Kolenbrander, Paul E.

    2009-01-01

    Formation of dental plaque is a developmental process involving initial and late colonizing species that form polymicrobial communities. Fusobacteria are the most numerous gram-negative bacteria in dental plaque, but they become prevalent after the initial commensal colonizers, such as streptococci and actinomyces, have established communities. The unusual ability of these bacteria to coaggregate with commensals, as well as pathogenic late colonizers, has been proposed to facilitate colonizat...

  7. Comprehensive discovery and characterization of small RNAs in Corynebacterium glutamicum ATCC 13032

    OpenAIRE

    Mentz, Almut; Neshat, Armin; Pfeifer, Katharina; Pühler, Alfred; Rückert, Christian; Kalinowski, Jörn

    2013-01-01

    Background Recent discoveries on bacterial transcriptomes gave evidence that small RNAs (sRNAs) have important regulatory roles in prokaryotic cells. Modern high-throughput sequencing approaches (RNA-Seq) enable the most detailed view on transcriptomes offering an unmatched comprehensiveness and single-base resolution. Whole transcriptome data obtained by RNA-Seq can be used to detect and characterize all transcript species, including small RNAs. Here, we describe an RNA-Seq approach for comp...

  8. Draft genome sequences of Streptococcus bovis strains ATCC 33317 and JB1

    Science.gov (United States)

    We report the draft genome sequences of Streptococcus bovis type strain ATTC 33317 (CVM42251) isolated from cow dung and strain JB1 (CVM42252) isolated from a cow rumen in 1977. Strains were subjected to Next Generation sequencing and the genome sizes are approximately 2 MB and 2.2 MB, respectively....

  9. Response of Bacillus cereus ATCC 14579 to challenges with sublethal concentrations of enterocin AS-48

    NARCIS (Netherlands)

    Grande Burgos, M.J.; Kovács, Á.T.; Mirończuk, A.M.; Abriouel, H.; Gálvez, A.; Kuipers, O.P.

    2009-01-01

    Background: Enterocin AS-48 is produced by Enterococcus faecalis S48 to compete with other bacteria in their environment. Due to its activity against various Gram positive and some Gram negative bacteria it has clear potential for use as a food preservative. Here, we studied the effect of enterocin

  10. 1,3-Propanediol production by Escherichia coli using genes from Citrobacter freundii atcc 8090.

    Science.gov (United States)

    Przystałowska, Hanna; Zeyland, Joanna; Kośmider, Alicja; Szalata, Marlena; Słomski, Ryszard; Lipiński, Daniel

    2015-01-01

    Compared with chemical synthesis, fermentation has the advantage of mass production at low cost, and has been used in the production of various industrial chemicals. As a valuable organic compound, 1,3-propanediol (1,3-PDO) has numerous applications in the production of polymers, lubricants, cosmetics and medicines. Here, conversion of glycerol (a renewable substrate and waste from biodiesel production) to 1,3-PDO by E. coli bacterial strain carrying altered glycerol metabolic pathway was investigated. Two gene constructs containing the 1,3-PDO operon from Citrobacter freundii (pCF1 and pCF2) were used to transform the bacteria. The pCF1 gene expression construct contained dhaBCE genes encoding the three subunits of glycerol dehydratase, dhaF encoding the large subunit of the glycerol dehydratase reactivation factor and dhaG encoding the small subunit of the glycerol dehydratase reactivating factor. The pCF2 gene expression construct contained the dhaT gene encoding the 1,3-propanediol dehydrogenase. Expression of the genes cloned in the above constructs was under regulation of the T7lac promoter. RT-PCR, SDS-PAGE analyses and functional tests confirmed that 1,3-PDO synthesis pathway genes were expressed at the RNA and protein levels, and worked flawlessly in the heterologous host. In a batch flask culture, in a short time applied just to identify the 1,3-PDO in a preliminary study, the recombinant E. coli bacteria produced 1.53 g/L of 1,3-PDO, using 21.2 g/L of glycerol in 72 h. In the Sartorius Biostat B Plus reactor, they produced 11.7 g/L of 1,3-PDO using 24.2 g/L of glycerol, attaining an efficiency of 0.58 [mol1,3-PDO/molglycerol].

  11. Proteomic evidences for rex regulation of metabolism in toxin-producing Bacillus cereus ATCC 14579.

    Science.gov (United States)

    Laouami, Sabrina; Clair, Géremy; Armengaud, Jean; Duport, Catherine

    2014-01-01

    The facultative anaerobe, Bacillus cereus, causes diarrheal diseases in humans. Its ability to deal with oxygen availability is recognized to be critical for pathogenesis. The B. cereus genome comprises a gene encoding a protein with high similarities to the redox regulator, Rex, which is a central regulator of anaerobic metabolism in Bacillus subtilis and other Gram-positive bacteria. Here, we showed that B. cereus rex is monocistronic and down-regulated in the absence of oxygen. The protein encoded by rex is an authentic Rex transcriptional factor since its DNA binding activity depends on the NADH/NAD+ ratio. Rex deletion compromised the ability of B. cereus to cope with external oxidative stress under anaerobiosis while increasing B. cereus resistance against such stress under aerobiosis. The deletion of rex affects anaerobic fermentative and aerobic respiratory metabolism of B. cereus by decreasing and increasing, respectively, the carbon flux through the NADH-recycling lactate pathway. We compared both the cellular proteome and exoproteome of the wild-type and Δrex cells using a high throughput shotgun label-free quantitation approach and identified proteins that are under control of Rex-mediated regulation. Proteomics data have been deposited to the ProteomeXchange with identifier PXD000886. The data suggest that Rex regulates both the cross-talk between metabolic pathways that produce NADH and NADPH and toxinogenesis, especially in oxic conditions.

  12. Proteomic evidences for rex regulation of metabolism in toxin-producing Bacillus cereus ATCC 14579.

    Directory of Open Access Journals (Sweden)

    Sabrina Laouami

    Full Text Available The facultative anaerobe, Bacillus cereus, causes diarrheal diseases in humans. Its ability to deal with oxygen availability is recognized to be critical for pathogenesis. The B. cereus genome comprises a gene encoding a protein with high similarities to the redox regulator, Rex, which is a central regulator of anaerobic metabolism in Bacillus subtilis and other Gram-positive bacteria. Here, we showed that B. cereus rex is monocistronic and down-regulated in the absence of oxygen. The protein encoded by rex is an authentic Rex transcriptional factor since its DNA binding activity depends on the NADH/NAD+ ratio. Rex deletion compromised the ability of B. cereus to cope with external oxidative stress under anaerobiosis while increasing B. cereus resistance against such stress under aerobiosis. The deletion of rex affects anaerobic fermentative and aerobic respiratory metabolism of B. cereus by decreasing and increasing, respectively, the carbon flux through the NADH-recycling lactate pathway. We compared both the cellular proteome and exoproteome of the wild-type and Δrex cells using a high throughput shotgun label-free quantitation approach and identified proteins that are under control of Rex-mediated regulation. Proteomics data have been deposited to the ProteomeXchange with identifier PXD000886. The data suggest that Rex regulates both the cross-talk between metabolic pathways that produce NADH and NADPH and toxinogenesis, especially in oxic conditions.

  13. Purification and enzymic properties of the fructosyltransferase of Streptococcus salivarius ATCC 25975.

    Science.gov (United States)

    Song, D D; Jacques, N A

    1999-01-01

    The recombinant fructosyltransferase (Ftf) of Streptococcus salivarius was expressed in Escherichia coli and purified to electrophoretic homogeneity after a combination of adsorption, ion-exchange and gel-filtration chromatography. The N-terminal signal sequence of the Ftf was removed by E. coli at the same site as in its natural host. The purified Ftf exhibited maximum activity at pH 6.0 and 37 degrees C, was activated by Ca2+, but inhibited by the metal ions Cu2+, Zn2+, Hg2+ and Fe3+. The enzyme catalysed the transfer of the fructosyl moiety of sucrose to a number of acceptors, including water, glucose and sucrose via a Ping Pong mechanism involving a fructosyl-enzyme intermediate. While this mechanism of catalysis is utilized by the levansucrases of Bacillus subtilis and Acetobacter diazotrophicus and the values of the kinetic constants for the three enzymes are similar, sucrose was a far more efficient fructosyl-acceptor for the Ftf of S. salivarius than for the two other enzymes. PMID:10393084

  14. Utilization of the cyanobacteria Anabaena sp. ATCC 33047 in CO2 removal processes.

    Science.gov (United States)

    González López, C V; Acién Fernández, F G; Fernández Sevilla, J M; Sánchez Fernández, J F; Cerón García, M C; Molina Grima, E

    2009-12-01

    In this paper the utilization of the cyanobacteria Anabaena sp. in carbon dioxide removal processes is evaluated. For this, continuous cultures of this strain were performed at different dilution rates; alternatives for the recovery of the organic matter produced being also studied. A maximum CO(2) fixation rate of 1.45 g CO(2) L(-1) day(-1) was measured experimentally, but it can be increased up to 3.0 g CO(2) L(-1) day(-1) outdoors. The CO(2) is mainly transformed into exopolysaccharides, biomass representing one third of the total organic matter produced. Organic matter can be recovered by sedimentation with efficiencies higher than 90%, the velocity of sedimentation being 2.10(-4) s(-1). The major compounds were carbohydrates and proteins with productivities of 0.70 and 0.12 g L(-1) day(-1), respectively. The behaviour of the cultures of Anabaena sp. has been modelized, also the characteristics parameters requested to design separation units being reported. Finally, to valorizate the organic matter as biofertilizers and biofuels is proposed.

  15. Draft Genome of Halomonas Species Strain GFAJ-1 (ATCC BAA-2256)

    OpenAIRE

    Phung, Le T.; Silver, Simon; Trimble, William L.; Gilbert, Jack A.

    2012-01-01

    Halomonas strain GFAJ-1 was reported in Science magazine to be a remarkable microbe for which there was “arsenate in macromolecules that normally contain phosphate, most notably nucleic acids.” The draft genome of the bacterium was determined (NCBI accession numbers AHBC01000001 through AHBC01000103). It appears to be a typical gamma proteobacterium.

  16. Characterization of the penicillin G acylase from Bacillus megaterium ATCC 14945

    Directory of Open Access Journals (Sweden)

    Vanessa Ribeiro de Souza

    2005-06-01

    Full Text Available The purpose of this work was to characterize the enzyme penicillin G acylase (PGA produced by Bacillus megaterium. Purification of the enzyme by ultra/diafiltration did not allow the detection of the PGA band by SDS-PAGE electrophoresis due to the high content of remaining proteins. However, using the DNA of the microorganism, it was possible to replicate the genes of the two B. megaterium PGA reported in literature, showing that the enzyme consisted of two sub-units, having 245 and 537 amino acids each and an average molecular mass of 26950 and 59070 Da, respectively. The parameters studied were: 1 the influence of temperature in the 25-60(0C range, 2 pH in the 5-10 range and 3 substrate concentration, this was tested to obtain results on the Penicillin G hydrolysis reaction rate, using the initial velocities approach. The maximum hydrolysis rate was obtained at 37ºC and pH 8.0. The Michaelis-Menten model fitted well, resulting in estimated Km and Vmax parameters values of 1.83 mM and 0.165*10-3 mmol/min/UI, respectively.O objetivo deste trabalho foi caracterizar a enzima penicilina G acilase (PGA produzida por Bacillus megaterium, uma importante enzima industrial que catalisa a hidrólise de penicilina G, para produção de antibióticos semi-sintéticos. Purificação da enzima por ultra/diafiltração não permitiu detectar a banda de PGA por eletroforese SDS-PAGE devido ao elevado conteúdo de outras proteínas remanescentes. Contudo, utilizando DNA do microrganismo que vem sendo estudado, foi possível amplificar os genes das duas sub-unidades de PGA previstas na literatura, mostrando que a enzima em estudo é também constituída de duas sub-unidades, 245 e 537 aminoácidos cada, com massas moleculares médias de 26950 e 59070 Da, respectivamente. Foram estudadas as influências da temperatura 25-60(0C, pH 5-10, e concentração do substrato na velocidade da reação de hidrólise da penicilina G. A temperatura e pH ótimos foram de 37(0C e 8,0, respectivamente. O modelo de Michaelis-Menten representou bem a cinética da reação, com valores de parâmetros estimados de 1,83 mM para Km e Vmáx= 0,165*10-3 mmol/min/UI.

  17. Inactivation of Escherichia coli ATCC 11775 in fresh produce using atmospheric pressure cold plasma

    Science.gov (United States)

    Bermudez-Aguirre, Daniela; Wemlinger, Erik; Barbosa-Canovas, Gustavo; Pedrow, Patrick; Garcia-Perez, Manuel

    2011-10-01

    Food-borne outbreaks are associated with the presence of pathogenic bacteria in food products such as fresh produce. One of the target microorganisms is Escherichia coli which exhibits resistance to being inactivated with conventional disinfection methods for vegetables. Atmospheric pressure cold plasma (APCP) was tested to disinfect three vegetables with challenge surfaces, lettuce, carrots and tomatoes. The produce was inoculated with the bacteria to reach an initial microbial concentration of 107 cfu/g. Vegetables were initially exposed to the APCP discharges from a needle array at 5.7 kV RMS in argon, processing times of 0.5, 3 and 5 min. Initial results indicate that microbial decontamination is effective on the lettuce (1.2 log reduction) when compared with other vegetables. To claim disinfection, a 3 log reduction or more is needed, which makes APCP treatment very promising technology for decontamination of produce. We propose that with method refinements full disinfection can be achieved using APCP.

  18. Determination of thermobacteriological parameters and size of Bacillus stearothermophilus ATCC 7953 spores

    Directory of Open Access Journals (Sweden)

    Marcos Fraiha

    2010-12-01

    Full Text Available In order to determine thermobacteriological parameters for B. stearothermophilus spores, they were diluted in a saline solution medium and in ground corn-soybean mix, distributed in TDT tube, and submitted to heat for a specific period of time. The D value (time to reduce 1 log cycle of microbial count under a certain temperature and z value (variation of temperature to cause 10-fold change in D value were estimated. To estimate their dimensions, the spores were visualized by using a scanning electron microscope. D121.1 ºC and z values for these spores, as determined in the saline solution, were 8.8 minutes and 12.8 ºC, respectively. D121,1 ºC and z values determined in the corn-soy mix were 14.2 minutes and 23.7 ºC, respectively. The micrographs indicated that the spores have homogeneous shape and size, with length and diameter of 2 and 1 µm, respectively. These results confirm that the spore is highly thermal-resistant, and it is a good biological indicator to evaluate the extrusion process as a feed sterilizer.

  19. Degradation of Arginine and Other Amino Acids by Eubacterium nodatum ATCC 33099

    OpenAIRE

    Uematsu, H.; Hoshino, E.

    2011-01-01

    The utilisation of a total of 20 amino acids by Eubacterium nodatum, a predominant asaccharolytic anaerobe isolated from human periodontal pockets, was studied. Washed cells of the microorganism produced substantial amounts of acetate, butyrate and ammonia from lysine, and butyrate and ammonia from arginine as main products under anaerobic conditions. They also produced a small amount of formate from histidine. Metabolic products were not detected from any of the other 17 amino acids. These r...

  20. Optimization of a Modified GS Medium for a Probiotic Strain (L. acidophilus ATCC4356

    Directory of Open Access Journals (Sweden)

    N. Pedram

    2014-09-01

    Full Text Available Probiotics are defined as living microorganisms with beneficial effects on the host. Probiotics, according to the least negative effect on the body, are considered as a good alternative to chemical drugs. Lactobacillus acidophilus is used as a probiotic that is able to reduce cholesterol level in the blood. The effect of various concentrations of carbon, nitrogen and phosphorus for enhancing the biomass production of Lactobacillus acidophilus was examined. The response surface methodology based on Box–Wilson CCD was applied to explore the optimal medium composition. Glucose, yeast extract, K2HPO4 and KH2PO4 were selected as dependent variables. All experiments were run at 37°C for 31h under stationary conditions. By solving the regression equation and analyzing the response surface carton, optimal concentrations of the components were determined as glucose (5-8.75 g.l-1, yeast extract (36.75-39 g.l-1, K2HPO4 (0.1-0.2125 g.l-1 and KH2PO4 (0.3925-0.7075 g.l-1. Validation experiment confirmed that the optimized medium was comparable to the MRS medium (the most common medium for Lactobacillus acidophilus strain in biomass production, having the advantages of economy andpracticality.

  1. Physiological roles for two periplasmic nitrate reductases in Rhodobacter sphaeroides 2.4.3 (ATCC 17025).

    Science.gov (United States)

    Hartsock, Angela; Shapleigh, James P

    2011-12-01

    The metabolically versatile purple bacterium Rhodobacter sphaeroides 2.4.3 is a denitrifier whose genome contains two periplasmic nitrate reductase-encoding gene clusters. This work demonstrates nonredundant physiological roles for these two enzymes. One cluster is expressed aerobically and repressed under low oxygen while the second is maximally expressed under low oxygen. Insertional inactivation of the aerobically expressed nitrate reductase eliminated aerobic nitrate reduction, but cells of this strain could still respire nitrate anaerobically. In contrast, when the anaerobic nitrate reductase was absent, aerobic nitrate reduction was detectable, but anaerobic nitrate reduction was impaired. The aerobic nitrate reductase was expressed but not utilized in liquid culture but was utilized during growth on solid medium. Growth on a variety of carbon sources, with the exception of malate, the most oxidized substrate used, resulted in nitrite production on solid medium. This is consistent with a role for the aerobic nitrate reductase in redox homeostasis. These results show that one of the nitrate reductases is specific for respiration and denitrification while the other likely plays a role in redox homeostasis during aerobic growth. PMID:21949073

  2. Investigation of the links between heterocyst and biohydrogen production by diazotrophic cyanobacterium A. variabilis ATCC 29413.

    Science.gov (United States)

    Salleh, Siti Fatihah; Kamaruddin, Azlina; Uzir, Mohamad Hekarl; Karim, Khairiah Abd; Mohamed, Abdul Rahman

    2016-03-01

    This work investigates the effect of heterocyst toward biohydrogen production by A. variabilis. The heterocyst frequency was artificially promoted by adding an amino acid analog, in this case DL-7-azatryptophan into the growth medium. The frequency of heterocyst differentiation was found to be proportional to the concentration of azatryptophan (0-25 µM) in the medium. Conversely, the growth and nitrogenase activity were gradually suppressed. In addition, there was also a distinct shortening of the cells filaments and detachment of heterocyst from the vegetative cells. Analysis on the hydrogen production performance revealed that both the frequency and distribution of heterocyst in the filaments affected the rate of hydrogen production. The highest hydrogen production rate and yield (41 µmol H2 mg chl a(-1) h(-1) and 97 mL H2 mg chl a(-1), respectively) were achieved by cells previously grown in 15 µM of azatryptophan with 14.5 % of heterocyst frequency. The existence of more isolated heterocyst has been shown to cause a relative loss in nitrogenase activity thus lowering the hydrogen production rate.

  3. Clostridium thermocellum ATCC27405 transcriptomic, metabolomic and proteomic profiles after ethanol stress

    Directory of Open Access Journals (Sweden)

    Yang Shihui

    2012-07-01

    Full Text Available Abstract Background Clostridium thermocellum is a candidate consolidated bioprocessing biocatalyst, which is a microorganism that expresses enzymes for both cellulose hydrolysis and its fermentation to produce fuels such as lignocellulosic ethanol. However, C. thermocellum is relatively sensitive to ethanol compared to ethanologenic microorganisms such as yeast and Zymomonas mobilis that are used in industrial fermentations but do not possess native enzymes for industrial cellulose hydrolysis. Results In this study, C. thermocellum was grown to mid-exponential phase and then treated with ethanol to a final concentration of 3.9 g/L to investigate its physiological and regulatory responses to ethanol stress. Samples were taken pre-shock and 2, 12, 30, 60, 120, and 240 min post-shock, and from untreated control fermentations for systems biology analyses. Cell growth was arrested by ethanol supplementation with intracellular accumulation of carbon sources such as cellobiose, and sugar phosphates, including fructose-6-phosphate and glucose-6-phosphate. The largest response of C. thermocellum to ethanol shock treatment was in genes and proteins related to nitrogen uptake and metabolism, which is likely important for redirecting the cells physiology to overcome inhibition and allow growth to resume. Conclusion This study suggests possible avenues for metabolic engineering and provides comprehensive, integrated systems biology datasets that will be useful for future metabolic modeling and strain development endeavors.

  4. 4-nitrocatechol as a probe of a Mn(II)-dependent extradiol-cleaving catechol dioxygenase (MndD): comparison with relevant Fe(II) and Mn(II) model complexes.

    Science.gov (United States)

    Reynolds, Mark F; Costas, Miquel; Ito, Masami; Jo, Du-Hwan; Tipton, A Alex; Whiting, Adam K; Que, Lawrence

    2003-02-01

    Mn(II)-dependent 3,4-dihydroxyphenylacetate 2,3-dioxygenase (MndD) is an extradiol-cleaving catechol dioxygenase from Arthrobacter globiformis that has 82% sequence identity to and cleaves the same substrate (3,4-dihydroxyphenylacetic acid) as Fe(II)-dependent 3,4-dihydroxyphenylacetate 2,3-dioxygenase (HPCD) from Brevibacterium fuscum. We have observed that MndD binds the chromophoric 4-nitrocatechol (4-NCH(2)) substrate as a dianion and cleaves it extremely slowly, in contrast to the Fe(II)-dependent enzymes which bind 4-NCH(2) mostly as a monoanion and cleave 4-NCH(2) 4-5 orders of magnitude faster. These results suggest that the monoanionic binding state of 4-NC is essential for extradiol cleavage. In order to address the differences in 4-NCH(2) binding to these enzymes, we synthesized and characterized the first mononuclear monoanionic and dianionic Mn(II)-(4-NC) model complexes as well as their Fe(II)-(4-NC) analogs. The structures of [(6-Me(2)-bpmcn)Fe(II)(4-NCH)](+), [(6-Me(3)-TPA)Mn(II)(DBCH)](+), and [(6-Me(2)-bpmcn)Mn(II)(4-NCH)](+) reveal that the monoanionic catecholate is bound in an asymmetric fashion (Delta r(metal-O(catecholate))=0.25-0.35 A), as found in the crystal structures of the E(.)S complexes of extradiol-cleaving catechol dioxygenases. Acid-base titrations of [(L)M(II)(4-NCH)](+) complexes in aprotic solvents show that the p K(a) of the second catecholate proton of 4-NCH bound to the metal center is half a p K(a) unit higher for the Mn(II) complexes than for the Fe(II) complexes. These results are in line with the Lewis acidities of the two divalent metal ions but are the opposite of the trend observed for 4-NCH(2) binding to the Mn(II)- and Fe(II)-catechol dioxygenases. These results suggest that the MndD active site decreases the second p K(a) of the bound 4-NCH(2) relative to the HPCD active site.

  5. Assessing the ecological long-term impact of wastewater irrigation on soil and water based on bioassays and chemical analyses.

    Science.gov (United States)

    Richter, Elisabeth; Hecht, Fabian; Schnellbacher, Nadine; Ternes, Thomas A; Wick, Arne; Wode, Florian; Coors, Anja

    2015-11-01

    The reuse of treated wastewater for irrigation and groundwater recharge can counteract water scarcity and reduce pollution of surface waters, but assessing its environmental risk should likewise consider effects associated to the soil. The present study therefore aimed at determining the impact of wastewater irrigation on the habitat quality of water after soil passage and of soil after percolation by applying bioassays and chemical analysis. Lab-scale columns of four different soils encompassing standard European soil and three field soils of varying characteristics and pre-contamination were continuously percolated with treated wastewater to simulate long-term irrigation. Wastewater and its percolates were tested for immobilization of Daphnia magna and growth inhibition of green algae (Pseudokirchneriella subcapitata) and water lentils (Lemna minor). The observed phytotoxicity of the treated wastewater was mostly reduced by soil passage, but in some percolates also increased for green algae. Chemical analysis covering an extensive set of wastewater-born organic pollutants demonstrated that many of them were considerably reduced by soil passage, particularly through peaty soils. Taken together, these results indicated that wastewater-born phytotoxic substances may be removed by soil passage, while existing soil pollutants (e.g. metals) may leach and impair percolate quality. Soils with and without wastewater irrigation were tested for growth of plants (Avena sativa, Brassica napus) and soil bacteria (Arthrobacter globiformis) and reproduction of collembolans (Folsomia candida) and oligochaetes (Enchytraeus crypticus, Eisenia fetida). The habitat quality of the standard and two field soils appeared to be deteriorated by wastewater percolation for at least one organism (enchytraeids, plants or bacteria), while for two pre-contaminated field soils it also was improved (for plants and/or enchytraeids). Wastewater percolation did not seem to raise soil concentrations

  6. Application of a new sediment contact test with Myriophyllum aquaticum and of the Aquatic Lemna test to assess the sediment quality of Lake Skadar

    Energy Technology Data Exchange (ETDEWEB)

    Stesevic, D.; Sundic, D.; Mijovic, S. [Montenegro Univ., Podgorica (ME). Faculty of Sciences; Feiler, U.; Heininger, P. [Federal Institute of Hydrology, Koblenz (Germany); Erdinger, L. [Heidelberg Univ. (Germany). Inst. of Hygiene; Seiler, T.B.; Hollert, H. [Heidelberg Univ. (Germany). Dept. of Zoology; RWTH Aachen (Germany). Inst. fuer Umweltforschung - Biologie V

    2007-10-15

    the sediment samples from Radus and Kamenik, whereas the aquatic Lemna test showed inhibition effects for the samples from Sterbeq, Plavnica and Kamice. Data obtained with the newly developed Danio rerio contact test and the Arthrobacter globiformis contact test confirmed the Myriophyllum results. Analyses of the heavy metal content in the sediments revealed low or moderate contamination levels. Correlation analyses between the content of heavy metals in the sediments and growth inhibition of Myriophyllum aquaticum showed a significant correlation between CR concentrations and growth inhibition. Comparable findings are available for a German river system. In contrast, no significant correlation between inhibition rates and concentration of metals could be observed with Lemna minor. (orig.)

  7. Comparative genomics of citric-acid-producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88

    DEFF Research Database (Denmark)

    Andersen, Mikael Rørdam; Salazar, Margarita Pena; Schaap, Peter J.;

    2011-01-01

    The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzyme...... and phylogenetics. Detailed lists of alleles were generated, and genotypic differences were observed to accumulate in metabolic pathways essential to acid production and protein synthesis. A transcriptome analysis supported up-regulation of genes associated with biosynthesis of amino acids that are abundant...

  8. 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

  9. Distinct substrate specificities of three glycoside hydrolase family 42 β-galactosidases from Bifidobacterium longum subsp. infantis ATCC 15697

    DEFF Research Database (Denmark)

    Viborg, Alexander Holm; Katayama, Takane; Abou Hachem, Maher;

    2014-01-01

    signatures of the three GH42 enzymes correlate to unique sequence motifs denoting specific clades in a GH42 phylogenetic tree providing novel insight into GH42 subspecificities. Overall, the data illustrate the metabolic adaptation of bifidobacteria to the β-galactoside-rich gut niche and emphasize...... enzymes enabling utilization of β-galactosides exerting prebiotic effects. However, insight into the specificity of individual GH42 enzymes with respect to substrate monosaccharide composition, glycosidic linkage and degree of polymerization is lagging. Kinetic analysis of natural and synthetic substrates...... (Galβ1-4Galβ1-4Gal). The specificity of Bga42C resembles that of Bga42B, but the activity was one order of magnitude lower. Based on enzyme kinetics, gene organization and phylogenetic analyses, Bga42C is proposed to act in the metabolism of arabinogalactan-derived oligosaccharides. The distinct kinetic...

  10. Influence of the initial ph of sugar cane juice on the production of levan by Zymomonas mobilis ATCC 31821

    Directory of Open Access Journals (Sweden)

    Marcia Sadae Tano

    2002-01-01

    Full Text Available The influence of initial pH of sugar cane juice in high concentration was investigated for levan production by Zymomonas mobilis. In the initial pH of medium of 5.4; 5.9 and 6..3 the levan concentration achieved 1.66; 2.54 and 3.83 g/L, respectively, in 48 hours of fermentation. The final levan concentration in the initial pH of 6.3 and 5.9 increased 130 and 53% when compared to concentration at initial pH 5.4 in the medium. The levan yield at pH 5.9 was 87.5% and at pH 6.3 was 162.5% superior in relation to that obtained at initial pH 5.4.

  11. Isolation and characterization of two cellulose morphology mutants of Gluconacetobacter hansenii ATCC23769 producing cellulose with lower crystallinity.

    Directory of Open Access Journals (Sweden)

    Ying Deng

    Full Text Available Gluconacetobacter hansenii, a Gram-negative bacterium, produces and secrets highly crystalline cellulose into growth medium, and has long been used as a model system for studying cellulose synthesis in higher plants. Cellulose synthesis involves the formation of β-1,4 glucan chains via the polymerization of glucose units by a multi-enzyme cellulose synthase complex (CSC. These glucan chains assemble into ordered structures including crystalline microfibrils. AcsA is the catalytic subunit of the cellulose synthase enzymes in the CSC, and AcsC is required for the secretion of cellulose. However, little is known about other proteins required for the assembly of crystalline cellulose. To address this question, we visually examined cellulose pellicles formed in growth media of 763 individual colonies of G. hansenii generated via Tn5 transposon insertion mutagenesis, and identified 85 that produced cellulose with altered morphologies. X-ray diffraction analysis of these 85 mutants identified two that produced cellulose with significantly lower crystallinity than wild type. The gene disrupted in one of these two mutants encoded a lysine decarboxylase and that in the other encoded an alanine racemase. Solid-state NMR analysis revealed that cellulose produced by these two mutants contained increased amounts of non-crystalline cellulose and monosaccharides associated with non-cellulosic polysaccharides as compared to the wild type. Monosaccharide analysis detected higher percentages of galactose and mannose in cellulose produced by both mutants. Field emission scanning electron microscopy showed that cellulose produced by the mutants was unevenly distributed, with some regions appearing to contain deposition of non-cellulosic polysaccharides; however, the width of the ribbon was comparable to that of normal cellulose. As both lysine decarboxylase and alanine racemase are required for the integrity of peptidoglycan, we propose a model for the role of peptidoglycan in the assembly of crystalline cellulose.

  12. Physiological Roles for Two Periplasmic Nitrate Reductases in Rhodobacter sphaeroides 2.4.3 (ATCC 17025)▿

    Science.gov (United States)

    Hartsock, Angela; Shapleigh, James P.

    2011-01-01

    The metabolically versatile purple bacterium Rhodobacter sphaeroides 2.4.3 is a denitrifier whose genome contains two periplasmic nitrate reductase-encoding gene clusters. This work demonstrates nonredundant physiological roles for these two enzymes. One cluster is expressed aerobically and repressed under low oxygen while the second is maximally expressed under low oxygen. Insertional inactivation of the aerobically expressed nitrate reductase eliminated aerobic nitrate reduction, but cells of this strain could still respire nitrate anaerobically. In contrast, when the anaerobic nitrate reductase was absent, aerobic nitrate reduction was detectable, but anaerobic nitrate reduction was impaired. The aerobic nitrate reductase was expressed but not utilized in liquid culture but was utilized during growth on solid medium. Growth on a variety of carbon sources, with the exception of malate, the most oxidized substrate used, resulted in nitrite production on solid medium. This is consistent with a role for the aerobic nitrate reductase in redox homeostasis. These results show that one of the nitrate reductases is specific for respiration and denitrification while the other likely plays a role in redox homeostasis during aerobic growth. PMID:21949073

  13. Role of C-Terminal Domains in Surface Attachment of the Fructosyltransferase of Streptococcus salivarius ATCC 25975

    Science.gov (United States)

    Rathsam, Catherine; Jacques, Nicholas A.

    1998-01-01

    The cell-associated β-d-fructosyltransferase of Streptococcus salivarius, which is devoid of the cell wall anchoring motif, LPXTG, is released on exposure to its substrate, sucrose. Deletions within the C terminus of the enzyme implicated both the hydrophobic and the proline-glycine-serine-threonine-rich wall-associated domain in stabilizing the enzyme on the cell surface. PMID:9829954

  14. Uptake and expulsion of sup 14 C-xylitol by xylitol-cultured Streptococcus mutans ATCC 25175 in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Soederling, E.; Pihlanto-Leppaelae, A. (Department of Biochemistry, Institute of Dentistry, University of Turku, Turku (Finland))

    1989-01-01

    The effect of successive cultivations in the presence of 6% xylitol on the uptake and expulsion of {sup 14}C-xylitol was studied using the cells of Streptococcus mutans 25175. Three sequential cultivations did not alter the growth inhibition percentage (approximately 50%) observed in the presence of 6% xylitol. The {sup 14}C-xylitol uptake experiments performed with growing and resting cells showed that both the uptake and the expulsion of xylitol were enhanced by xylitolculturing. Both xylitol-cultured and resting control cells contained only one major labeled compound which was identified as {sup 14}C-xylitol 5-phosphate. The label subsequently was expelled from the cells as {sup 14}C-xylitol. These results indicate that S. mutans possesses an intracellular xylitol cycle and this cycle is regulated by adding xylitol to the growth medium. (author).

  15. Silver Nanoparticles: Biosynthesis Using an ATCC Reference Strain of Pseudomonas aeruginosa and Activity as Broad Spectrum Clinical Antibacterial Agents

    Science.gov (United States)

    Quinteros, Melisa A.; Aiassa Martínez, Ivana M.; Dalmasso, Pablo R.; Páez, Paulina L.

    2016-01-01

    Currently, the biosynthesis of silver-based nanomaterials attracts enormous attention owing to the documented antimicrobial properties of these ones. This study reports the extracellular biosynthesis of silver nanoparticles (Ag-NPs) using a Pseudomonas aeruginosa strain from a reference culture collection. A greenish culture supernatant of P. aeruginosa incubated at 37°C with a silver nitrate solution for 24 h changed to a yellowish brown color, indicating the formation of Ag-NPs, which was confirmed by UV-vis spectroscopy, transmission electron microscopy, and X-ray diffraction. TEM analysis showed spherical and pseudospherical nanoparticles with a distributed size mainly between 25 and 45 nm, and the XRD pattern revealed the crystalline nature of Ag-NPs. Also it provides an evaluation of the antimicrobial activity of the biosynthesized Ag-NPs against human pathogenic and opportunistic microorganisms, namely, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Proteus mirabilis, Acinetobacter baumannii, Escherichia coli, P. aeruginosa, and Klebsiella pneumonia. Ag-NPs were found to be bioactive at picomolar concentration levels showing bactericidal effects against both Gram-positive and Gram-negative bacterial strains. This work demonstrates the first helpful use of biosynthesized Ag-NPs as broad spectrum bactericidal agents for clinical strains of pathogenic multidrug-resistant bacteria such as methicillin-resistant S. aureus, A. baumannii, and E. coli. In addition, these Ag-NPs showed negligible cytotoxic effect in human neutrophils suggesting low toxicity to the host. PMID:27340405

  16. Improved activity and pH stability of E-coli ATCC 11105 penicillin acylase by error-prone PCR

    NARCIS (Netherlands)

    Balci, Huseyin; Ozturk, Merve Tuzlakoglu; Pijning, Tjaard; Ozturk, Saliha Issever; Gumusel, Fusun

    2014-01-01

    Penicillin G acylase is the key enzyme used in the industrial production of β-lactam antibiotics. This enzyme hydrolyzes penicillin G and related β-lactam antibiotics releasing 6-aminopenicillanic acid, which is an intermediate in the production of semisynthetic penicillins. To improve the enzymatic

  17. BUTYRIC ACID AND HYDROGEN PRODUCTION BY C CLOSTRIDIUM TYROBUTYRICUM ATCC 25755 AND M MUTANTS. (R829479C016)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  18. Influence of the initial ph of sugar cane juice on the production of levan by Zymomonas mobilis ATCC 31821

    OpenAIRE

    Marcia Sadae Tano; João Batista Buzato

    2002-01-01

    The influence of initial pH of sugar cane juice in high concentration was investigated for levan production by Zymomonas mobilis. In the initial pH of medium of 5.4; 5.9 and 6..3 the levan concentration achieved 1.66; 2.54 and 3.83 g/L, respectively, in 48 hours of fermentation. The final levan concentration in the initial pH of 6.3 and 5.9 increased 130 and 53% when compared to concentration at initial pH 5.4 in the medium. The levan yield at pH 5.9 was 87.5% and at pH 6.3 was 162.5% superio...

  19. Uptake and expulsion of 14C-xylitol by xylitol-cultured Streptococcus mutans ATCC 25175 in vitro

    International Nuclear Information System (INIS)

    The effect of successive cultivations in the presence of 6% xylitol on the uptake and expulsion of 14C-xylitol was studied using the cells of Streptococcus mutans 25175. Three sequential cultivations did not alter the growth inhibition percentage (approximately 50%) observed in the presence of 6% xylitol. The 14C-xylitol uptake experiments performed with growing and resting cells showed that both the uptake and the expulsion of xylitol were enhanced by xylitolculturing. Both xylitol-cultured and resting control cells contained only one major labeled compound which was identified as 14C-xylitol 5-phosphate. The label subsequently was expelled from the cells as 14C-xylitol. These results indicate that S. mutans possesses an intracellular xylitol cycle and this cycle is regulated by adding xylitol to the growth medium. (author)

  20. Complete genome sequence of the facultatively chemolithoautotrophic and methylotrophic alpha Proteobacterium Starkeya novella type strain (ATCC 8093T)

    Energy Technology Data Exchange (ETDEWEB)

    Kappler, Ulrike [University of Queensland, The, Brisbane, Queensland, Australia; Davenport, Karen W. [Los Alamos National Laboratory (LANL); Beatson, Scott [University of Queensland, The, Brisbane, Queensland, Australia; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Berry, Kerrie W. [U.S. Department of Energy, Joint Genome Institute; Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Hammon, Nancy [U.S. Department of Energy, Joint Genome Institute; Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Richardson, P M [U.S. Department of Energy, Joint Genome Institute; Bruce, David [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Tapia, Roxanne [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Chang, Yun-Juan [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Goker, Markus [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Klenk, Hans-Peter [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute

    2012-01-01

    Starkeya novella (Starkey 1934) Kelly et al. 2000 is a member of the family Xanthobacteraceae in the order Rhizobiales , which is thus far poorly characterized at the genome level. Cultures from this spe- cies are most interesting due to their facultatively chemolithoautotrophic lifestyle, which allows them to both consume carbon dioxide and to produce it. This feature makes S. novella an interesting model or- ganism for studying the genomic basis of regulatory networks required for the switch between con- sumption and production of carbon dioxide, a key component of the global carbon cycle. In addition, S. novella is of interest for its ability to grow on various inorganic sulfur compounds and several C1- compounds such as methanol. Besides Azorhizobium caulinodans, S. novella is only the second spe- cies in the family Xanthobacteraceae with a completely sequenced genome of a type strain. The cur- rent taxonomic classification of this group is in significant conflict with the 16S rRNA data. The ge- nomic data indicate that the physiological capabilities of the organism might have been underestimat- ed. The 4,765,023 bp long chromosome with its 4,511 protein-coding and 52 RNA genes was se- quenced as part of the DOE Joint Genome Institute Community Sequencing Program (CSP) 2008.

  1. Lime application for the efficient production of nutraceutical glucooligosaccharides from Leuconostoc mesenteroides NRRL B-742 (ATCC13146).

    Science.gov (United States)

    Moon, Young Hwan; Madsen, Lee; Chung, Chang-Ho; Kim, Doman; Day, Donal F

    2015-02-01

    We have previously demonstrated the production of glucooligosaccharides via a fermentation of sucrose with Leuconostoc mesenteroides NRRL B-742 using sodium hydroxide (NaOH) to control the pH. Because NaOH is expensive, we sought to minimize the cost of our process by substituting hydrated lime and saccharate of lime (lime sucrate) in its place. The yield of glucooligosaccharides using either 5 % lime (41.4 ± 0.5 g/100 g) or 5 % lime sucrate (40.0 ± 1.4 g/100 g) were both similar to the NaOH control (42.4 ± 1.5 g/100 g). Based on this, it appears that the cost associated with pH control in our process can be reduced by a factor of approximately 2.4 using lime instead of NaOH. Because our chromatographic stage is based on a Ca(2+)-form resin to separate glucooligosaccharides, the use of lime not only negates the need for costly de-salting via ion-exchange (elimination of two ion-exchange sections) prior to separation, but also greatly reduces the resin regeneration cost. PMID:25533635

  2. Impact of impurities in biodiesel-derived crude glycerol on the fermentation by Clostridium pasteurianum ATCC 6013

    Energy Technology Data Exchange (ETDEWEB)

    Venkataramanan, Keerthi P.; Boatman, Judy J.; Taconi, Katherine A. [Alabama Univ., Huntsville, AL (United States). Dept. of Chemical and Materials Engineering; Kurniawan, Yogi; Bothun, Geoffrey D. [Rhode Island Univ., Kingston, RI (United States). Dept. of Chemical Engineering; Scholz, Carmen [Alabama Univ., Huntsville, AL (United States). Dept. of Chemistry

    2012-02-15

    During the production of biodiesel, crude glycerol is produced as a byproduct at 10% (w/w). Clostridium pasteurianum has the inherent potential to grow on glycerol and produce 1,3-propanediol and butanol as the major products. Growth and product yields on crude glycerol were reported to be slower and lower, respectively, in comparison to the results obtained from pure glycerol. In this study, we analyzed the effect of each impurity present in the biodiesel-derived crude glycerol on the growth and metabolism of glycerol by C. pasteurianum. The crude glycerol contains methanol, salts (in the form of potassium chloride or sulfate), and fatty acids that were not transesterified. Salt and methanol were found to have no negative effects on the growth and metabolism of the bacteria on glycerol. The fatty acid with a higher degree of unsaturation, linoleic acid, was found to have strong inhibitory effect on the utilization of glycerol by the bacteria. The fatty acid with lower or no degrees of unsaturation such as stearic and oleic acid were found to be less detrimental to substrate utilization. The removal of fatty acids from crude glycerol by acid precipitation resulted in a fermentation behavior that is comparable to the one on pure glycerol. These results show that the fatty acids in the crude glycerol have a negative effect by directly affecting the utilization of glycerol as the carbon source, and hence their removal from crude glycerol is an essential step towards the utilization of crude glycerol. (orig.)

  3. Isolation and characterization of two cellulose morphology mutants of Gluconacetobacter hansenii ATCC23769 producing cellulose with lower crystallinity.

    Science.gov (United States)

    Deng, Ying; Nagachar, Nivedita; Fang, Lin; Luan, Xin; Catchmark, Jeffrey M; Tien, Ming; Kao, Teh-hui

    2015-01-01

    Gluconacetobacter hansenii, a Gram-negative bacterium, produces and secrets highly crystalline cellulose into growth medium, and has long been used as a model system for studying cellulose synthesis in higher plants. Cellulose synthesis involves the formation of β-1,4 glucan chains via the polymerization of glucose units by a multi-enzyme cellulose synthase complex (CSC). These glucan chains assemble into ordered structures including crystalline microfibrils. AcsA is the catalytic subunit of the cellulose synthase enzymes in the CSC, and AcsC is required for the secretion of cellulose. However, little is known about other proteins required for the assembly of crystalline cellulose. To address this question, we visually examined cellulose pellicles formed in growth media of 763 individual colonies of G. hansenii generated via Tn5 transposon insertion mutagenesis, and identified 85 that produced cellulose with altered morphologies. X-ray diffraction analysis of these 85 mutants identified two that produced cellulose with significantly lower crystallinity than wild type. The gene disrupted in one of these two mutants encoded a lysine decarboxylase and that in the other encoded an alanine racemase. Solid-state NMR analysis revealed that cellulose produced by these two mutants contained increased amounts of non-crystalline cellulose and monosaccharides associated with non-cellulosic polysaccharides as compared to the wild type. Monosaccharide analysis detected higher percentages of galactose and mannose in cellulose produced by both mutants. Field emission scanning electron microscopy showed that cellulose produced by the mutants was unevenly distributed, with some regions appearing to contain deposition of non-cellulosic polysaccharides; however, the width of the ribbon was comparable to that of normal cellulose. As both lysine decarboxylase and alanine racemase are required for the integrity of peptidoglycan, we propose a model for the role of peptidoglycan in the assembly of crystalline cellulose. PMID:25790428

  4. Genome Sequence of Virgibacillus pantothenticus DSM 26T (ATCC 14576), a Mesophilic and Halotolerant Bacterium Isolated from Soil.

    Science.gov (United States)

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

    2015-01-01

    Virgibacillus pantothenticus DSM 26(T) is a Gram-positive, spore-forming, aerobic, mesophilic, and halotolerant bacterium. Here, we report its 4.76-Mb draft genome sequence, which is the first genome information of V. pantothenticus and will promote biological research and biotechnological application for the species. PMID:26383648

  5. Genome Sequence of Virgibacillus pantothenticus DSM 26T (ATCC 14576), a Mesophilic and Halotolerant Bacterium Isolated from Soil

    OpenAIRE

    Wang, Jie-ping; Liu, Bo; Liu, Guo-hong; Chen, De-ju; Zhu, Yu-jing; Chen, Zheng; Che, Jian-mei

    2015-01-01

    Virgibacillus pantothenticus DSM 26T is a Gram-positive, spore-forming, aerobic, mesophilic, and halotolerant bacterium. Here, we report its 4.76-Mb draft genome sequence, which is the first genome information of V. pantothenticus and will promote biological research and biotechnological application for the species.

  6. The effect of rhizosphere on growth of Sphingomonas chlorophenolica ATCC 39723 during pentachlorophenol (PCP biodegradation in batch culture and soil

    Directory of Open Access Journals (Sweden)

    Ken Killhan

    2006-12-01

    Full Text Available Studies on the influence of the rhizosphere on the growth of Sphingomonas chlorophenolica during Pentacholophenol (PCP degradation in batch culture and in soil were carried out. In batch culture, a basal minimal medium with or without rhizosphere exudates extracted from winter wheat was used. In soil systems, degradation experiments were performed in the presence and absence of plants. Measurements of PCP concentrations were made using high performance liquid chromatography analysis (HPLC. Bacterial analyses of S. chlorophenolica were carried out by plating on MSM medium. The results showed that the rhizosphere exudates stimulated the growth of the cells of S. chlorophenolica at concentrations of 50 and 80mg kg dry wt soil –1 as well as stimulating the ability of S. chlorophenolica to degrade PCP at a concentration of 80mg Kg dry wt soil -1. In addition, pentachlorophenol had an adverse effect on the growth of S. chlorophenolica. The introduction of S.chlorophenolica into the loamy soil with plants showed a faster degradation when compared to the inoculated soil without plants. There was a significant increase of S. chlorophenolica in the roots in comparison to those in the soil. This study showed that the presence of the inoculum S. chlorophenolica enhanced the PCP degradation in a loamy soil and it indicates the potential for a treatment process under a appropriate environmental conditions such as there present in soil systems.

  7. Transcriptional regulation of metabolic pathways, alternative respiration and enterotoxin genes in anaerobic growth of Bacillus cereus ATCC 14579

    NARCIS (Netherlands)

    Voort, van der M.; Abee, T.

    2009-01-01

    Aims: To assess genes specifically activated during anaerobic growth that are involved in metabolism and pathogenesis of the foodborne pathogen Bacillus cereus. Methods and Results: Growth under anaerobic conditions in Brain Heart Infusion (BHI) broth revealed a reduced growth rate and lower yield a

  8. Protein (Cyanobacteria): 435829 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available 3658 Cyanothece sp. ATCC 51142 MSCSIQIPALSMTSVTPPNVNQQTIELAPSYNIPIILILMAIATLLIQPWVSLPLALFGLFLLLQTVTIRLQFTATALDVYRSDQRIRSFPYSEWQNWKIFWQPIPILFYFKEVNSIHFLPIIFDPQTLNVCLERFCNFDKMEEMG ...

  9. Protein (Cyanobacteria): 34863 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available anothece sp. ATCC 51142 MAKRVQVVLNESINKLGKDGDLVEVAPGYARNYLLPQKLATLATPGILKQVEQRREKERQRLLAEKQQAEGQKTALETIKRFTIRKEVGEGEAIFGTVTTSEVAEAIQAATNQEVDRKGITVPDINKTGFYEATVKLHPEVTATIEIQVAPL ...

  10. Characterization of three antifungal calcite-forming bacteria, Arthrobacter nicotianae KNUC2100, Bacillus thuringiensis KNUC2103, and Stenotrophomonas maltophilia KNUC2106, derived from the Korean islands, Dokdo and their application on mortar.

    Science.gov (United States)

    Park, Jong-Myong; Park, Sung-Jin; Ghim, Sa-Youl

    2013-09-28

    Crack remediation on the surface of cement mortar using microbiological calcium carbonate (CaCO3) precipitation (MICP) has been investigated as a microbial sealing agent on construction materials. However, MICP research has never acknowledged the antifungal properties of calcite-forming bacteria (CFB). Since fungal colonization on concrete surfaces can trigger biodeterioration processes, fungi on concrete buildings have to be prevented. Therefore, to develop a microbial sealing agent that has antifungal properties to remediate cement cracks without deteriorative fungal colonization, we introduced an antifungal CFB isolated from oceanic islands (Dokdo islands, territory of South Korea, located at the edge of the East Sea in Korea.). The isolation of CFB was done using B4 or urea-CaCl2 media. Furthermore, antifungal assays were done using the pairing culture and disk diffusion methods. Five isolated CFB showed CaCO3 precipitation and antifungal activities against deteriorative fungal strains. Subsequently, five candidate bacteria were identified using 16S rDNA sequence analysis. Crack remediation, fungi growth inhibition, and water permeability reduction of antifungal CFB-treated cement surfaces were tested. All antifungal CFB showed crack remediation abilities, but only three strains (KNUC2100, 2103, and 2106) reduced the water permeability. Furthermore, these three strains showed fungi growth inhibition. This paper is the first application research of CFB that have antifungal activity, for an eco-friendly improvement of construction materials.

  11. Effect of pulsed light treatments on the growth and resistance behavior of Listeria monocytogenes 10403S, Listeria innocua, and Escherichia coli ATCC 25922 in a liquid substrate.

    Science.gov (United States)

    Uesugi, Aaron R; Hsu, Lillian; Moraru, Carmen I

    2013-03-01

    Pulsed light (PL) treatment can effectively inactivate a large proportion of contaminating bacteria on surfaces and in clear solutions. An important issue that needs to be investigated is whether repeated exposure to PL treatment causes any changes to the growth and resistance behavior of the bacteria surviving the treatment. To test this, three challenge microorganisms were used: Listeria monocytogenes, Listeria innocua, and Escherichia coli. Cells of the challenge bacteria were treated with either low or high PL doses. Survivors of the PL treatment were enumerated, isolated, regrown, and exposed again to PL treatment. PL inactivation curves were generated for the survivors of each exposure cycle (as well as controls) to examine possible differences induced by repeated treatments. Growth curves of L. monocytogenes, L. innocua, and E. coli isolates recovered from exposure to either 1.1 or 10.1 J/cm(2) were not significantly different from the growth curves of untreated cells. Reduction levels of up to 4 and up to 6 log CFU were obtained after exposure to 1.1 and 10.1 J/cm(2), respectively, both for the controls and the repeatedly treated and recovered isolates. These results show that PL did not significantly change the growth kinetics or resistance to PL of the target microorganisms after up to 10 exposures. These findings have significance for the practical application of PL treatment, as they indicate that this technology does not select for microorganisms with increased resistance.

  12. β-galactosidase Production by Aspergillus niger ATCC 9142 Using Inexpensive Substrates in Solid-State Fermentation: Optimization by Orthogonal Arrays Design

    Science.gov (United States)

    Kazemi, Samaneh; Khayati, Gholam; Faezi-Ghasemi, Mohammad

    2016-01-01

    Background: Enzymatic hydrolysis of lactose is one of the most important biotechnological processes in the food industry, which is accomplished by enzyme β-galactosidase (β-gal, β-D-galactoside galactohydrolase, EC 3.2.1.23), trivial called lactase. Orthogonal arrays design is an appropriate option for the optimization of biotechnological processes for the production of microbial enzymes. Methods: Design of experimental (DOE) methodology using Taguchi orthogonal array (OA) was employed to screen the most significant levels of parameters, including the solid substrates (wheat straw, rice straw, and peanut pod), the carbon/nitrogen (C/N) ratios, the incubation time, and the inducer. The level of β-gal production was measured by a photometric enzyme activity assay using the artificial substrate ortho-Nitrophenyl-β-D-galactopyranoside. Results: The results showed that C/N ratio (0.2% [w/v], incubation time (144 hour), and solid substrate (wheat straw) were the best conditions determined by the design of experiments using the Taguchi approach. Conclusion: Our finding showed that the use of rice straw and peanut pod, as solid-state substrates, led to 2.041-folds increase in the production of the enzyme, as compared to rice straw. In addition, the presence of an inducer did not have any significant impact on the enzyme production levels.

  13. Cloning and sequencing of the kedarcidin biosynthetic gene cluster from Streptoalloteichus sp. ATCC 53650 revealing new insights into biosynthesis of the enediyne family of antitumor antibiotics†

    OpenAIRE

    Lohman, Jeremy R.; Huang, Sheng-Xiong; Horsman, Geoffrey P.; Dilfer, Paul E.; Huang, Tingting; Chen, Yihua; Wendt-Pienkowski, Evelyn; Shen, Ben

    2013-01-01

    Enediyne natural product biosynthesis is characterized by a convergence of multiple pathways, generating unique peripheral moieties that are appended onto the distinctive enediyne core. Kedarcidin (KED) possesses two unique peripheral moieties, a (R)-2-aza-3-chloro-β-tyrosine and an iso-propoxy-bearing 2-naphthonate moiety, as well as two deoxysugars. The appendage pattern of these peripheral moieties to the enediyne core in KED differs from the other enediynes studied to date with respect to...

  14. Two-component signal transduction system CBO0787/CBO0786 represses transcription from botulinum neurotoxin promoters in Clostridium botulinum ATCC 3502.

    Directory of Open Access Journals (Sweden)

    Zhen Zhang

    2013-03-01

    Full Text Available Blocking neurotransmission, botulinum neurotoxin is the most poisonous biological substance known to mankind. Despite its infamy as the scourge of the food industry, the neurotoxin is increasingly used as a pharmaceutical to treat an expanding range of muscle disorders. Whilst neurotoxin expression by the spore-forming bacterium Clostridium botulinum appears tightly regulated, to date only positive regulatory elements, such as the alternative sigma factor BotR, have been implicated in this control. The identification of negative regulators has proven to be elusive. Here, we show that the two-component signal transduction system CBO0787/CBO0786 negatively regulates botulinum neurotoxin expression. Single insertional inactivation of cbo0787 encoding a sensor histidine kinase, or of cbo0786 encoding a response regulator, resulted in significantly elevated neurotoxin gene expression levels and increased neurotoxin production. Recombinant CBO0786 regulator was shown to bind to the conserved -10 site of the core promoters of the ha and ntnh-botA operons, which encode the toxin structural and accessory proteins. Increasing concentration of CBO0786 inhibited BotR-directed transcription from the ha and ntnh-botA promoters, demonstrating direct transcriptional repression of the ha and ntnh-botA operons by CBO0786. Thus, we propose that CBO0786 represses neurotoxin gene expression by blocking BotR-directed transcription from the neurotoxin promoters. This is the first evidence of a negative regulator controlling botulinum neurotoxin production. Understanding the neurotoxin regulatory mechanisms is a major target of the food and pharmaceutical industries alike.

  15. Impact of environmental factors on viability and stability and high pressure pretreatment on stress tolerance of Lactobacillus rhamnosus GG (ATCC 53103) during spray drying

    OpenAIRE

    Ananta, Edwin

    2005-01-01

    Diese Arbeit befasst sich in erster Linie mit der Evaluierung der Anwendbarkeit von Sprühtrocknung als eine alternative Methode zur Herstellung von Probiotika enthaltenden Trockenpräparaten. Dazu wurden optimale Trocknungsbedingungen ermittelt und die verwendeten Schutzmedien auf thermo-physikalische Beschaffenheit und Wechselwirkung mit einem synthetischen Membransystem untersucht. Eine Trocknungstemperatur von 80 °C wurde als ein annehmbarer Kompromiss ermittelt. Bei dieser Temperatur ließ ...

  16. Bacillus cereus ATCC 14579 RpoN (Sigma 54) is a Pleiotropic Regulator of Growth, Carbohydrate, Metabolism, Motility, Biofilm Formation and Toxin Production

    NARCIS (Netherlands)

    Hayrapetyan, H.; Tempelaars, M.H.; Nierop Groot, M.N.; Abee, T.

    2015-01-01

    Sigma 54 is a transcriptional regulator predicted to play a role in physical interaction of bacteria with their environment, including virulence and biofilm formation. In order to study the role of Sigma 54 in Bacillus cereus, a comparative transcriptome and phenotypic study was performed using B. c

  17. Significance of CO2 donor on the production of succinic acid by Actinobacillus succinogenes ATCC 55618

    OpenAIRE

    Zou Wei; Zhu Li-Wen; Li Hong-Mei; Tang Ya-Jie

    2011-01-01

    Abstract Background Succinic acid is a building-block chemical which could be used as the precursor of many industrial products. The dissolved CO2 concentration in the fermentation broth could strongly regulate the metabolic flux of carbon and the activity of phosphoenolpyruvate (PEP) carboxykinase, which are the important committed steps for the biosynthesis of succinic acid by Actinobacillus succinogenes. Previous reports showed that succinic acid production could be promoted by regulating ...

  18. Investigation of xylose and arabinose metabolism in Clostridium acetobutylicum ATCC 824 and Clostridium saccharobutylicum NCP 262.

    OpenAIRE

    Lesiak, Justyna Maria

    2015-01-01

    Xylulose kinase (xylB) and arabinose kinase (araK) mutants of Clostridium acetobutylicum and C. saccharobutylicum were investigated and compared to wild type strains, regarding their abilities to metabolize xylose, arabinose and glucose. To create mutants in C. saccharobutylicum, an effective triparental mating system for DNA transfer was developed, and additionally restrictase mutants were constructed and characterized. Furthermore, growth and gene expression of C. acetobutylicum in continuo...

  19. Enumeration of sublethally injured Escherichia coli O157:H7 ATCC 43895 and Escherichia coli strain B-41560 using selective agar overlays versus commercial methods.

    Science.gov (United States)

    Smith, Amanda R; Ellison, Alysha L; Robinson, Amanda L; Drake, Maryanne; McDowell, Susan A; Mitchell, James K; Gerard, Patrick D; Heckler, Rachel A; McKillip, John L

    2013-04-01

    Quality control procedures during food processing may involve direct inoculation of food samples onto appropriate selective media for subsequent enumeration. However, sublethally injured bacteria often fail to grow, enabling them to evade detection and intervention measures and ultimately threaten the health of consumers. This study compares traditional selective and nonselective agar-based overlays versus two commercial systems (Petrifilm and Easygel) for recovery of injured E. coli B-41560 and O157:H7 strains. Bacteria were propagated in tryptic soy broth (TSB), ground beef slurry, and infant milk formula to a density of 10(6) to 10(8) CFU/ml and then were stressed for 6 min either in lactic acid (pH 4.5) or heat shocked for 3 min at 60°C. Samples were pour plated in basal layers of either tryptic soy agar (TSA), sorbitol MacConkey agar (SMAC), or violet red bile agar (VRB) and were resuscitated for 4 h prior to addition of agar overlays. Other stressed bacteria were plated directly onto Petrifilm and Easygel. Results indicate that selective and nonselective agar overlays recovered significantly higher numbers (greater than 1 log) of acid- and heat-injured E. coli O157:H7 from TSB, ground beef, and infant milk formula compared with direct plating onto selective media, Petrifilm, or Easygel, while no significant differences among these media combinations were observed for stressed E. coli B-41560. Nonstressed bacteria from TSB and ground beef were also recovered at densities significantly higher in nonselective TSA-TSA and in VRB-VRB and SMAC-SMAC compared with Petrifilm and Easygel. These data underscore the need to implement food safety measures that address sublethally injured pathogens such as E. coli O157:H7 in order to avoid underestimation of true densities for target pathogens.

  20. A β-glucosidase from Oenococcus oeni ATCC BAA-1163 with potential for aroma release in wine: Cloning and expression in E. coli

    OpenAIRE

    Michlmayr, Herbert; Schümann, Christina; Wurbs, Phillip; Barreira Braz da Silva, Nuno M.; Rogl, Veronika; Kulbe, Klaus D.; del Hierro, Andrés M

    2010-01-01

    Lactic acid bacteria (LAB) are responsible for olfactory changes in wine during malolactic fermentation (MLF). A side characteristic of MLF is the release of grape derived aroma compounds from their glycosylated precursors by β-glycosidase activities of these bacteria. Apart from Oenococcus oeni, which is regarded as the most promising species for MLF, glycosidic activities have also been observed in wine related members of the genera Lactobacillus and Pediococcus. Nevertheless, information o...