Complete Genome Sequence of pAP13, a Large Linear Plasmid of a Brevibacterium Strain Isolated from a Saline Lake at 4,200 Meters above Sea Level in Argentina

pAP13 is an 89 kb-linear plasmid hosted by Brevibacterium sp. Ap13, an actinobacterium isolated from the feces of a flamingo from an extremely high-altitude lake in Argentina. Because of the ecological importance of the genus Brevibacterium, the absolute lack of information concerning Brevibacterium linear plasmids, as well as the as the possible ecological significance of this unusual plasmid, pAP13 was completely sequenced, including the inversely oriented termini.Fil: Dib, Julian Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Schuldes, Jörg. Georg August Universität; AlemaniaFil: Thürner, Andrea. Georg August Universität; AlemaniaFil: Farias, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Daniel, Rolf. Georg August Universität; AlemaniaFil: Meinhardt, Friedhelm. No especifíca

First Complete Sequence of a Giant Linear Plasmid from a Micrococcus Strain Isolated from an Extremely High-Altitude Lake

Micrococcus sp. V7, an actinobacterial strain adapted to the extreme conditions of the Laguna Vilama, an extremely high-altitude (4,600mabove sea level) lake in the Argentinian Puna, was found to carry the giant linear plasmid pLMV7. We determined its sequence (92,815 bp) as a prerequisite to the investigation of its role for survival in such a harsh environment.Fil: Dib, Julian Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Schuldes, Jörg. Georg August Universität Göttingen; AndorraFil: Thürner, Andrea. Georg August Universität Göttingen; AndorraFil: Farias, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Daniel, Rolf. Georg August Universität Göttingen; AndorraFil: Meinhardt, Friedhelm. No especifíca

Complete genome sequence of Staphylococcus aureus 6850, a highly cytotoxic and clinically virulent Methicillin-sensitive strain with distant relatedness to prototype strains

Staphylococcus aureus is a frequent human commensal bacterium and pathogen. Here we report the complete genome sequence of strain 6850 (spa type t185; sequence type 50 [ST50]), a highly cytotoxic and clinically virulent methicillin-sensitive strain from a patient with complicated S. aureus bacteremia associated with osteomyelitis and septic arthritis

Nueva esterasa tolerante a los solventes orgánicos aislada por metagenómica: ideas sobre la clasificación de las esterasas/lipasas

In order to isolate novel organic solvent-tolerant (OST) lipases, a metagenomic library was built using DNA derived from a temperate forest soil sample. A two-step activity-based screening allowed the isolation of a lipolytic clone active in the presence of organic solvents. Sequencing of the plasmid pRBest recovered from the positive clone revealed the presence of a putative lipase/esterase encoding gene. The deduced amino acid sequence (RBest1) contains the conserved lipolytic enzyme signature and is related to the previously described OST lipase from Lysinibacillus sphaericus 205y, which is the sole studied prokaryotic enzyme belonging to the 4.4 / hydrolase subgroup (abH04.04). Both in vivo and in vitro studies of the substrate specificity of RBest1, using triacylglycerols or nitrophenyl-esters, respectively, revealed that the enzyme is highly specific for butyrate (C4) compounds, behaving as an esterase rather than a lipase. The RBest1 esterase was purified and biochemically characterized. The optimal esterase activity was observed at pH 6.5 and at temperatures ranging from 38 to 45 °C. Enzymatic activity, determined by hydrolysis of p nitrophenyl esters, was found to be affected by the presence of different miscible and non-miscible organic solvents, and salts. Noteworthy, RBest1 remains significantly active at high ionic strength. These findings suggest that RBest1 possesses the ability of OST enzymes to molecular adaptation in the presence of organic compounds and resistance of halophilic proteins.Con el fin de aislar nuevas variantes de lipasas tolerantes a solventes orgánicos (OST), se construyó una librería metagenómica a partir de ADN obtenido de una muestra de suelo de bosque templado. A través de un monitoreo en dos etapas, basado en la detección de actividades, se aisló un clon con actividad lipolítica en presencia de solventes orgánicos. La secuenciación del plásmido pRBest recuperado del clon positivo reveló la presencia de un gen codificante de una hipotética lipasa/esterasa. La secuencia deducida de amino ácidos (RBest1) contiene los motivos conservados de enzimas lipolíticas y está relacionada con la lipasa OST previamente descrita de Lysinibacillus sphaericus 205y, que es la única enzima procariota estudiada perteneciente al subgrupo 4.4 de α/β hidrolasas (abH4.04). Estudios in vivo e in vitro sobre la especificidad de sustratos de RBest1, utilizando triacil-gliceroles o p-nitrofenil-ésteres, respectivamente, revelaron que la enzima es altamente específica para compuestos butíricos (C4 ), comportándose como una esterasa y no como una lipasa. La esterasa RBest1 fue purificada y caracterizada bioquímicamente. La actividad óptima de esterasa fue observada a pH 6,5 y las temperaturas óptimas fueron entre 38 y 45 °C. Se estableció que la actividad enzimática, determinada por hidrólisis de p-nitrofenil ésteres, es afectada en presencia de diferentes solventes orgánicos miscibles y no miscibles, y también sales. Notoriamente, RBest1 permanece significativamente activa a elevadas fuerzas iónicas. Estos hallazgos sugieren que RBest1 posee la capacidad de las enzimas OST de la adaptación molecular en presencia de compuestos orgánicos, así como la resistencia de las proteínas halófilasFil: Berlemont, Renaud. Universite de Liege; BélgicaFil: Spee, Olivier. Universite de Liege; BélgicaFil: Delsaute, Maud. Universite de Liege; BélgicaFil: Lara, Yannick. Universite de Liege; BélgicaFil: Schuldes, Jörg. Universitat of Gottingen; AlemaniaFil: Simon, Carola. Universitat of Gottingen; AlemaniaFil: Power, Pablo. Universite de Liege; Bélgica. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Daniel, Rolf. Universitat of Gottingen; AlemaniaFil: Galleni, Moreno. Universite de Liege; Bélgic

Novel organic solvent-tolerant esterase isolated by metagenomics: insights into the lipase/esterase classification

in order to isolate novel organic solvent-tolerant (oSt) lipases, a metagenomic library was built using dna derived from a temperate forest soil sample. a two-step activity-based screening allowed the isolation of a lipolytic clone active in the presence of organic solvents. Sequencing of the plasmid pRBest recovered from the positive clone revealed the presence of a putative lipase/esterase encoding gene. the deduced amino acid sequence (RBest1) contains the conserved lipolytic enzyme signature and is related to the previously described oSt lipase from Lysinibacillus sphaericus 205y, which is the sole studied prokaryotic enzyme belonging to the 4.4 a/b hydrolase subgroup (abh04.04). Both in vivo and in vitro studies of the substrate specificity of RBest1, using triacylglycerols or nitrophenyl-esters, respectively, revealed that the enzyme is highly specific for butyrate (c4) compounds, behaving as an esterase rather than a lipase. the RBest1 esterase was purified and biochemically characterized. the optimal esterase activity was observed at ph 6.5 and at temperatures ranging from 38 to 45 °c. enzymatic activity, determined by hydrolysis of p‐nitrophenyl esters, was found to be affected by the presence of different miscible and non-miscible organic solvents, and salts. noteworthy, RBest1 remains significantly active at high ionic strength. these findings suggest that RBest1 possesses the ability of oSt enzymes to molecular adaptation in the presence of organic compounds and resistance of halophilic proteins

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

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 quinaldine, respectively

Complete Genome Sequence of the Linear Plasmid pJD12 Hosted by Micrococcus sp. D12, Isolated from a High-Altitude Volcanic Lake in Argentina

The linear plasmid pDJ12 from Micrococcus D12, isolated from the high-altitude volcanic Diamante Lake in the northwest of Argentina, was completely sequenced and annotated. It is noteworthy that the element is probably conjugative and harbors genes potentially instrumental in coping with stress conditions that prevail in such an extreme environmentFil: Dib, Julian Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Planta Piloto de Procesos Industriales Microbiológicos (i); Argentina. Universidad Nacional de Tucumán; ArgentinaFil: Angelov, Angel. Technische Universitat Munchen; AlemaniaFil: Liebl, Wolfang. Technische Universitat Munchen; AlemaniaFil: Döbber, Johannes. Westfalische Wilhelms Universitat; AlemaniaFil: Voget, Sonja. Georg-August University; AlemaniaFil: Schuldes, Jörg. Georg-August University; AlemaniaFil: Gorriti, Marta Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Planta Piloto de Procesos Industriales Microbiológicos (i); ArgentinaFil: Farias, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Planta Piloto de Procesos Industriales Microbiológicos (i); ArgentinaFil: Meinhardt, Friedhelm. Westfalische Wilhelms Universitat; AlemaniaFil: Daniele, Rolf. Georg-August University; Alemani

Genome sequence analyses of two isolates from the recent Escherichia coli outbreak in Germany reveal the emergence of a new pathotype: Entero-Aggregative-Haemorrhagic Escherichia coli (EAHEC)

The genome sequences of two Escherichia coli O104:H4 strains derived from two different patients of the 2011 German E. coli outbreak were determined. The two analyzed strains were designated E. coli GOS1 and GOS2 (German outbreak strain). Both isolates comprise one chromosome of approximately 5.31 Mbp and two putative plasmids. Comparisons of the 5,217 (GOS1) and 5,224 (GOS2) predicted protein-encoding genes with various E. coli strains, and a multilocus sequence typing analysis revealed that the isolates were most similar to the entero-aggregative E. coli (EAEC) strain 55989. In addition, one of the putative plasmids of the outbreak strain is similar to pAA-type plasmids of EAEC strains, which contain aggregative adhesion fimbrial operons. The second putative plasmid harbors genes for extended-spectrum β-lactamases. This type of plasmid is widely distributed in pathogenic E. coli strains. A significant difference of the E. coli GOS1 and GOS2 genomes to those of EAEC strains is the presence of a prophage encoding the Shiga toxin, which is characteristic for enterohemorrhagic E. coli (EHEC) strains. The unique combination of genomic features of the German outbreak strain, containing characteristics from pathotypes EAEC and EHEC, suggested that it represents a new pathotype Entero-Aggregative-Haemorrhagic Escherichiacoli (EAHEC)

Characterization of the microbial diversity of extreme habitats in the Kamchatka-region

Die im äußersten Nordosten von Russland gelegene Halbinsel Kamtschatka ist überzogen von heißen Quellen, Geysiren und Vulkanen. Proben aus diesen extremen Habitaten sind für die Entdeckung neuartiger Biokatalysatoren interessant. Als Ausgangsmaterial für diese Arbeit dienten Sedimente, Wasser und Bodenproben im Temperaturbereich von 50 bis 91 °C und pH-Bereichen von 2 bis 3,5 und 8,3 bis 9,5. Durch die Methode des mechanischen Bead-beating wurde die metagenomische DNA isoliert. Diese diente als Grundlage zur Konstruktion von Metagenombanken und schließlich zur Identifizierung neuartiger Biokatalysatoren und Wirkstoffe. Im Rahmen der auf Aktivität basierenden Durchmusterung der Genbanken konnten vier verschiedene Lipasen/Esterasen, zwei Proteasen und eine Amylase identifiziert und teilweise charakterisiert werden. Die isolierte metagenomische DNA diente schließlich auch zur Untersuchung der mikrobiellen Diversität. Mittels 16S rRNA-Genanalysen wurden 508 16S rRNA-Gensequenzen generiert, analysiert und in phylogenetische Stammbäume integriert. Es konnten 16S rRNA-Gensequenzen der Phyla Proteobacteria, Bacteroidetes, Chlorobi, Acidobacteria, Firmicutes, Nitrospirae, Planctomyces, Actinobacteria, Chloroflexi, Gemmatimonadetes, Cyanobacteria, Aquificae, Thermotogae, Crenarchaeota und Euryarchaeota sowie der Candidates Divisions OP8, OP10, OP11, TM6 und BRC1 identifiziert werden