VAAM-Jahrestagung 2012 18.–21. März in Tübingen

164The study results indicated that some strains of iron bacteria were veryeffective to remove diclofenac under axenic condition. Among the 18tested strains, 4 strains showed removal efficiency above 90%.[1] Ternes TA, 1998. Occurrence of drugs in German sewage treatment plants and rivers. Water Res 32:3245-3260[2] Heberer T, Reddersen K, Mechlinski A, 2002. From municipal sewage to drinking water: fate andremoval of pharmaceutical residues in the aquatic environment in urban areas. Water Sci. Technol. 46:81-88[3] Ashton D, Hilton M, Thomas KV, 2004. Investigating the environmental transport of humanpharmaceuticals to streams in the United Kingdom. Sci Total Environ 333:167-184[4] Kim SD, Cho J, Kim IS, Vanderford BJ, Snyder SA, 2007. Occurrence and removal of pharmaceuticalsand endocrine disruptors in South Korean surface, drinking , and waste waters. Water Res 41(5):1013-1021[5] Zhang YJ, Geissen SU, Gal C, 2008b. Carbamazepine and diclofenac: removal in wastewater treatmentplants and occurrence in water bodies. Chemosphere 73: 1151-1161[6] Monteiro SC, Boxall ABA, 2010. Occurrence and fate of human pharmaceuticals in the environment.Rev Environ Contam Toxicol 202:53-154OTP123Fluorescence microscopical analysis of PHB granuleassociated proteins (PGAPs) in Ralstonia eutropha H16D. Rais*, D. Pfeiffer*, D. JendrossekInstitut für Mikrobiologie, Universität Stuttgart, Stuttgart, GermanyRalstonia eutropha H16 has become the model organism for studyingmetabolism of poly(3-hydroxybutyrate) (PHB), an importantbiodegradable biopolymer [1]. Despite > 2 decades of intense research onPHB metabolism new PHB granule-associated proteins were recentlydiscovered using a two hybrid screening approach [2]. Meanwhile, at least19 proteins are known that are important for biosynthesis, maintenance andintracellular mobilization of PHB in R. eutropha. These are: acetoacetyl-CoA-thiolase (PhaA) and acetoacetyl-CoA reductase (PhaB) that arenecessary for synthesis of the PHB monomer (3-hydroxybutyryl-CoA),five phasinproteins (PhaPs) that include major constituents of the granulessurface layer, 9 PHB depolymerases (PhaZs) (two of which are oligomerhydrolases), PHB synthase (PhaC), regulator PhaR and recently discoveredPhaM that ensures equal distribution of PHB granules during cell division[3]. Many of the above-mentioned proteins presumably are attached to thePHB granules surface layer. However, only for PhaC, PhaR, PhaP1,PhaP5, PhaZa1 and PhaM data are published that confirminvivoattachment of these PGAPs. In this study we determined subcellularlocalization of all currently known 5 phasin proteins (PhaP1-5) undercondition permissive and restrictive for PHB accumulation. N- and C-terminal fusions of the respective phasin protein with eYfp wereconstructed and the respective fusions cloned on a braod host rangeplasmid were conjugatively transfered to R. eutropha. All fusions wereexpressed in the wild type H16, in strain PHB-4 (a chemically inducedmutant with a nonsense mutation in PhaC) and in a chromosomal phaCmutant. Similar fusions were constructed for all putative PHBdepolymerases. The results for the depolymerases will be presented in aseparated poster (A. Sznajder et al.).[1] Reinecke, F., Steinbüchel, A. (2009). J. Mol. Microbiol. Biotechnol. 16:91-108[2] Pfeiffer D., Jendrossek D. (2011). Microbiology. 157:2795-807.[3] Pfeiffer D., Wahl A., Jendrossek D. (2011). Mol Microbiol.82:936-51.OTP124Elucidating the CRISPR-Cas-System of Methanosarcina mazeiGö1D. Jäger, K. Weidenbach, L. Nickel, R. Schmitz-Streit*Christian-Albrechts-Universität Kiel, Institut für Allgemeine Mikrobiologie,Kiel, GermanyMethanosarcina mazei strain Gö1 (M. mazei) belongs to themethylotrophic methanogens of the order Methanosarcinales, which havethe most versatile substrate spectrum within the methanogenic archaeacontributing significantly to the production of the green house gas (Rogers& Whitman, 1991; Thauer, 1998).The genome annotation published in 2002 (Deppenmeier et al., 2002) didnot include the information on potential CRISPR loci in archaeal modelorganism. Our recent investigations however identified the presence of twomain CRISPR loci in M. mazei. As characteristic for CRISPR loci, both ofthem contain a conserved direct repeat of 37 nucleotides in length. Thefirst CRISPR locus, containing 47 direct repeats with spacers, is flanked bya Cas type I-B system, whereas the second locus (containing 81 directrepeats) is flanked by a polycistronic operon encoding a RAMP module ofCAS proteins (type III-B). Interestingly, based on sequence homology ofalready known Cas6 proteins, none of the loci obviously encode for themajor endoribonuclease of crRNA maturation. Here we present theidentification of potential M. mazei Cas6 orthologs. The biochemicalcharacterization of the protein(s) will be presented and discussed.Deppenmeier, U., Johann, A., Hartsch, T., Merkl, R., Schmitz, R. A., Martinez-Arias, R., Henne, A., et al.(2002). The genome of Methanosarcina mazei: evidence for lateral gene transfer between bacteria andarchaea.J Mol Microbiol Biotechnol,4(4), 453-461.Rogers, J. E., & Whitman eds., W. B. (1991). Microbial production and consumption of greenhouse gases:methane, nitrogen oxides and halomethanes.ASM Press, Washington DC.Thauer, R. K. (1998). Biochemistry of methanogenesis: a tribute to Marjory Stephenson. 1998 MarjoryStephenson Prize Lecture.Microbiology,144 ( Pt 9, 2377-2406.OTP125SMC is recruited to oriC by ParB and promotes chromosomesegregation in Streptococcus pneumoniae and Bacillus subtilisA. Minnen* 1 , L. Attaiech 2 , M. Thon 2 , *F. Bürmann 1 , J.-W. Veening 2 ,S. Gruber 11 Max Planck Institute of Biochemistry, Chromosome Organization andDynamics, Martinsried, Germany2 Groningen Biomolecular Sciences and Biotechnology Institute, Groningen,NetherlandsReliable segregation of replicated chromosomes is a prerequisite formaintaining genomic integrity. Multi-protein complexes formed by theStructural Maintenance of Chromosomes (SMC) proteins are essentialplayers for performing this task both in mitosis and meiosis, as well asduring the bacterial cell cycle.SMC proteins are highly conserved in all domains of life. Most bacteriaexpress a single SMC that is associated with the kleisin ScpA and ScpBprotein to form a complex called "bacterial condensin". In many bacterialspecies condensin is indispensable for proper chromosome condensationand segregation.We found that condensins of Bacillus subtilis and the human pathogenStreptococcus pneumoniae promote segregation of replicatedchromosomes and are recruited to parS sites at the origin of replication bythe sequence specific DNA binding protein ParB. This targetingmechanism seems to be conserved at least among gram-positive bacteriaand can be reconstituted in a heterologous expression system.OTP126Bacterial cytoskeletal element MreB forms dynamic actin-likefilaments in live cells and in vitroH.J. Defeu Soufo* 1 , C. Reimold 1 , H. Breddermann 1 , P. von Ohlshausen 2 ,A. Rohrbach 2 , P.L. Graumann 11 Albert-Ludwigs-Universität, Institut für Mikrobiologie , Freiburg, Germany2 Albert-Ludwigs-Universität, Institut für Mikrosystemtechnik, Freiburg,GermanyMreB protein is an essential component of the cell shape generationsystem and additionally affects many subcellular positioning processes inbacteria. MreB has a three dimensional structure that is highly similar tothat of actin and forms filamentous structures in vitro. However, it has stillbeen a matter of dispute if actin and MreB have arisen through divergentor convergent evolution. It has recently been proposed that the activity ofMreB does not depend on the formation of extended filaments and that theprotein forms patch like structures rather than dynamic filaments in vivo.Using super resolution microscopy (S-TIRF) with a resolution of 100 nm,we provide evidence that MreB forms filaments in live Bacillus subtilisbacteria, which can extend at a rate of 65 nm/s and mostly have a length inbetween half and full turns around the cell periphery. Filaments display asurprisingly variable degree of orientations, from circumferential tohelical, can fuse and split, and show extension dynamics that are affectedthrough a point mutation within the ATPase motif. FRAP experimentsreveal very fast exchange rates consistent with rapid filament turnover.MreB and its three paralogs Mbl and MreBH also form polymers in vitro,dependent on ATP and magnesium. Our results demonstrate that MreBforms extended filamentous structures that are able to confer long rangeinteractions with membrane proteins, which can be circumferential as wellas helical. Given that any polymer has an inherent bending stiffness, andthat MreB filaments are mostly longer than a half turn around the cellperiphery, filaments may exert a mechanical force against the membranethat can lead to local transfer of energy against the wall, possiblyfacilitating the incorporation of new peptidoglycan strands into the existingwall polymer. Our data further support the notion that MreB and actin havehad a common ancestor whose function was already based on dynamicfilament extension/retraction reactions.OTP127Analysis of the complete genome of Janthinobacterium sp. HH01reveals a homoserine lactone-independent regulation of theviolacein biosynthesis genesC. Hornung* 1 , A. Poehlein 2 , M. Schmidt 1 , M. Blokesch 3 , R. Daniel 2 ,W. Streit 11 Universität Hamburg, Biozentrum Klein Flottbek, Mikrobiologie undBiotechnologie, Hamburg, Germany2 Georg-August-University of Göttigen, Göttingen Genomics Laboratory,Institute of Microbiology and Genetics, Göttingen, Germany3 Polytechnique Fédérale de Lausanne (EPFL), Laboratory of MolecularMicrobiology, Global Health Institute, Lausanne, Switzerland, SwitzerlandThe gram-negative -proteobacterium Janthinobacterium sp. HH01 wasrecently isolated from an aquatic environment. Janthinobacteria formbeneficial biofilms on the skin of amphibia and are involved in preventingfungal growth [1,2]. HH01 grows well in a wide temperature rangebetween 4 and 17 °C and produces violacein in stationary growth phase.BIOspektrum | Tagungsband 2012