biofilm centre annual report 2012 - uni-due.de · 2014-01-23 · aquatic biotechnology mainly...
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Biofilm Centre
Annual Report 2012
Aquatic Microbiology
Aquatic Biotechnology
Molecular Enzyme Technology & Biochemistry
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Biofilm Centre
The Biofilm Centre is composed of three groups: Aquatic Microbiology, Aquatic Biotechnology and Molecular Enzyme Technology & Biochemistry. It is part of the Faculty of Chemistry and exists since 2001 at the University of Duisburg-Essen. By the end of 2010, the Biofilm Centre moved completely from three different locations at the campus Duisburg into one location at the campus Essen. That was a major effort, but now we have settled and make more and more use of the new, improved situation. An example is the project on “Stress response on biofilms of the thermoacidophilic archaeon Sulfolobus acidocaldarius”, in which all three groups of the Biofilm Centre are involved. The success of the Biofilm Centre is also reflected in the total amount of funding of the projects running in 2012, which was 5.25 Mio €!
The Biofilm Centre comprised in 2012 a staff of 49, with three professors, one Research Group Leader, 3 laboratory heads, 6 postdocs, 29 Ph. D. students, 6 Technicians, 2 Secretaries.
The focus of the Aquatic Microbiology lies in fundamental biofilm research and on the role of biofilms as a habitat for pathogens with Jost Wingender as leader of the “Research Group Pathogens in Biofilms”. Hans-Curt Flemming is affiliated to the IWW Water Centre as a Member of the Board of Directors. During the last three years, a DFG founded project on the role of plankton as a habitat for hygienically relevant microorganisms has been almost completed. Furthermore, five large projects, funded by the Ministry of Research and Technology on biofilm management in drinking water installations, on the efficacy of silver nanoparticles in implants, and on the desiccation resistance of microorganisms under space conditions have been granted and run.
In 2011, Prof. Flemming was appointed as Visiting Professor at the Singapore Center for Life Sciences and Engineering at the National Technical University of Singapore. In 2010 and 2011, he was invited for 9 keynote presentations on national and international conferences.
In 2012, a new research project was granted with the title „Safe Ruhr“. It is dedicated to the occurrence of hygienically relevant microorganisms in River Ruhr water in terms of recreational use and of optimization of drinking water treatment. The project has a volume of 780.000 € and lasts until 2015
Also in 2012, Prof. Flemming was appointed as the chairman of the BioCluster, a joint specialist group of the International Water Association (IWA) and the International Society of Microbial Ecology. This cluster is an approach to bring water specialists in closer contact with the latest developments in microbial ecology.
Aquatic Biotechnology mainly focuses on fundamentals and applications of biological leaching processes, biocorrosion and the prevention of acid mine drainage. The group deals with microbial surface processes such as the bioleaching of metal sulfides and the microbial influenced corrosion (MIC) of concrete and steel. Consequently, our scientific expertise is the microbiology of the iron, manganese, and sulfur cycles, which include moderate and extreme acidophilic iron and sulfur compound-oxidizing species (e. g. of the genera Acidithiobacillus and Leptospirillum) and manganese ion-oxidizing as well as sulfate-reducing bacteria.
In 2012, a project on “Urban Mining” was granted by the Ministery of Research and Technology. It is dedicated to the extraction of valuable elements from brown coal ashes by bioleaching. Furthermore, a cooperative research project with the Ruhr-Universität Bochum indicates that switching from planktonic lifestyle to Biofilm lifestyle causes in acidophilic bacteria the activation of about 100 genes and the inactivation of 50 genes (accepted for publication in "Proteomics")
Prof. Sand was appointed as a Guest Professor at the Central South University in Changsha, Hunan, China, from 2012 bis 2016
Molecular Enzyme Technology and Biochemistry concentrates on elucidating archaeal carbohydrate metabolism and stress response including transcription and transcription
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regulation. Beside classical biochemical, molecular biological and genetic approaches in collaboration with (inter)national partners state of the art high-throughput approaches (e.g. transcriptomics, proteomics, metabolomics and modeling) are used to gain a system level understanding (systems biology).
The general aim of the group of Prof. Siebers (MEB) is to unravel carbohydrate metabolism and its regulation, stress response as well as biofilm formation (e.g. synthesis of extracellular polymeric substances in archaeal model organisms (Siebers et al. 2012, Ulas et al. 2012, Esser at al. 2012). Based on fundamental research one major focus are biotechnological applications of extremozymes, extremophiles (metabolic engineering) as well as catalytic biofilms (Koerdt et al. 2012).
A new research project was granted in 2012 by the Ministery of Research and Technology (2013-2015): e:Bio – Innovationswettbewerb Systembiologie (Verbundprojekt Modul II, Transfer), „Applied Sulfolobus Systems Biology; Exploiting the hot archaeal metabolic potential for Biotechnology - SulfoSYSBIOTEC.
Under the leadership of Dr. Siebers 10 partners, amongst others Sigma Aldrich, will be engaged to unravel the central carbohydrate metabolism of the thermoacidophilic Archaeon Sulfolobus solfataricus (78-80°C, pH 2-3). The aim is to provide new enzymes “extremozymes” for the production of fine chemicals and process optimization (e.g. cellulose degradation in the area of renewable resources). (Total budget 2.550.940 €, Siebers Group: 623.666 €)
With an overall funding of ~5.2 Mio € for projects currently in progress, it ranks among the top groups of our university. The funding is provided by the German Research Foundation, the Ministry for Research and Education, the Ministry for Environment, the German Environmental Foundation the Ministry for Economy, the European Union and from industry.
For the Biofilm Centre:
Bettina Siebers Hans-Curt Flemming Wolfgang Sand
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Table of Contents
Biofilm Centre Staff ............................................................................................................. 6 Projects Aquatic Microbiolgy
Safe Ruhr – Bathing Water and Drinking Water for the Ruhr Area ........................................ 8 (Pl: Hans-Curt Flemming, Jost Wingender, Martin Strathmann)
Recognition and countermeasures to transiently non-culturable pathogens in drinking water installations ........................................................................................................................... 9 (Pl: Hans-Curt Flemming, Jost Wingender)
Nanosilver particles - mechanisms of action and study of possible interaction with tissue, cells and molecules. Definition of their relevant potential for intolerance...............................10 (Pl: Hans-Curt Flemming, Jost Wingender)
Microbial ochering in technical systems ................................................................................11 (Pl: Hans-Curt Flemming, Jost Wingender)
Dynaklim (“Dynamic adaptation of regional planning and developmental processes to the impact of climate change in the Emscher-Lippe region (Ruhrgebiet)” ...................................12 (Pl: Hans-Curt Flemming, Jost Wingender)
BOSS – Biofilm Organisms Surfing Space. ..........................................................................13 (Pl: Hans-Curt Flemming, Jost Wingender)
Proteins, polysaccharides and nucleic acids as components of the extracellular polymeric substances of drinking-water biofilms ...................................................................................14 (Pl: Hans-Curt Flemming, Jost Wingender)
Freshwater plankton as a habitat for hygienically relevant microorganisms ..........................15 (Pl: Hans-Curt Flemming, Jost Wingender)
Advanced Technologies for Water Resource Management (ATWARM) Marie-Curie project 16 (Pl: Hans-Curt Flemming, Thorsten Schmidt)
Membrane biofouling: hydraulic resistance of biofilms ..........................................................17 (Pl: Hans-Curt Flemming, Hans Vrouvenvelder)
Projects Aquatic Biotechnology
Iron- and sulfur- oxidizing microorganisms in the Lusatian- and Central German lignite mining districts .................................................................................................................................18 (Pl: Wolfgang Sand)
Attachment to and dissolution of metal sulfides by moderately thermophilic archaeon Ferroplasma .........................................................................................................................19 (Pl: Wolfgang Sand)
Urban mining – Recovery of metals from lignite ashes .........................................................20 (Pl: Wolfgang Sand)
Microcalorimetry – a Measuring Technique for the Detection of Biomining ...........................21 (Pl: Wolfgang Sand)
Nutrition optimization of thermophilic and halophilic phototrophic microorganisms ...............22 (Pl: Wolfgang Sand)
BIOCOR Initial Training Network (ITN) on biocorrosion ........................................................23 (Pl: Wolfgang Sand)
Biogenic sulfuric acid corrosion of different materials ...........................................................24 (Pl: Wolfgang Sand)
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Physiological and Molecular processes involved in Biofilm formation in Bioleaching bacteria .............................................................................................................................................26 (Pl: Wolfgang Sand)
Projects Molecular Enzyme Technology & Biochemistry
HotZyme - Systematic screening for novel hydrolases from hot environments .....................27 (Pl: Bettina Siebers)
Stress response on Biofilms of the thermoacidophilic Archaeon Sulfolobus acidocaldarius ..28 (Pl: Bettina Siebers)
ExpresSYS - A Platform of Novel Microbial Expression Systems for Industrially Relevant Genes ..................................................................................................................................29 (Pl: Bettina Siebers)
Study of the microbiological effect of silver nanoparticles and silver doped CaP nanoparticles. Synthesis, characterization and biological effects. .........................................30 (Pl: Bettina Siebers)
Hot biofilms: Biofilm formation and EPS synthesis of the thermoacidophilic Archaeon Sulfolobus acidocaldarius .....................................................................................................31 (Pl: Bettina Siebers)
Hot signal transduction in Sulfolobus: Regulatory protein phosphorylation in thermoacidophilic Archaea ...................................................................................................32 (Pl: Bettina Siebers)
Trehalose as stress response in (hyper-)thermophilic Archaea ............................................33 (Pl: Bettina Siebers)
Functional analysis of multiple general transcription factors in the crenarchaeal model organism Sulfolobus acidocaldarius .....................................................................................34 (Pl: Bettina Siebers)
Bachelor Theses accomplished 2012 within or supervised by the Biofilm Centre ........35
Master Theses accomplished 2012 within or supervised by the Biofilm Centre ...........38
Publications of Biofilm Centre in 2010..............................................................................40
Articles 2011 .......................................................................................................................41
Articles 2012 .......................................................................................................................42
Articles 2013 .......................................................................................................................43
Invited Lectures ..................................................................................................................45
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Biofilm Centre Staff
Birgit Manger Secretary Claudia Johänning Secretary Aquatic Microbiology Hans-Curt Flemming Head Jost Wingender Head of laboratory
Leader of “Research Group on Pathogens in Biofilms” Giacomo Bertini Ph. D. student Gaby Czeczor Technician Astrid Dannehl Technician Barbara Derricks Ph. D. student Zenyta Dwidjosiswojo Ph. D. student Jan Frösler Ph. D. student Marina Horstkott Ph. D. student Anika Marko Technician Witold Michalowski Postdoc Alexa Pillen Ph. D. student Conny Rosengarten Technician Miriam Tewes Ph. D. student Janine Wagner Ph. D. student Aquatic Biotechnology Wolfgang Sand Head Tilman Gehrke Head of laboratory Cindy-Jane Africa Ph. D. student Sören Bellenberg Ph. D. student Laura Castro Ph. D. student Iaryna Datsenko Research employee Bianca Florian Ph. D. student Claudia Janosch Ph. D. student Inga Kirstein Ph. D. student Beate Krok Ph. D. student Andrzej Kuklinski Ph. D. student Quian Li Ph. D. student Xiarong Liu Visiting professor Nanni Noel Ph. D. student Jürgen Schrötz Ph. D. student Christian Thyssen Ph. D. student Petra Wahl Technician Agata Wikiel Ph. D. student Mario Vera Postdoc Rujong Zhang Ph. D. student Molecular Enzyme Technology & Biochemistry Bettina Siebers Head Christopher Bräsen Head of laboratory Jens Benninghoff Ph. D. student Sabine Dietl Technician
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Dominik Esser Ph. D. student Anna Hagemann Ph. D. student Silke Jachlewski Ph. D. student Verena Kallnik Postdoc Thomas Knura Technician Julia Kort Ph. D. student Theresa Kouril Postdoc Kohei Matsubara Ph. D. student Bernadette Rauch Ph. D. student Britta Tjaden Postdoc
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Safe Ruhr – Bathing Water and Drinking Water for the Ruhr Area
Collaborative Project - Partners: IWW Water Centre, RWW Rheinisch-Westfälische Wasserwerksgesellschaft mbH, University of Duisburg-Essen (Aquatic Ecology; Institute for Communication Science; Institute for Sociology), Ruhrverband, University of Bonn, University of Bochum, RWTH Aachen, Karlsruhe Institute of Technology (KIT), aquatune GmbH, Xylem Water Solutions Coordinator: Dr. Martin Strathmann (IWW) Funded by: Federal Ministry of Education and Research (BMBF); Network: Risk Management of Emerging Compounds and Pathogens in the Water Cycle (RiSKWa) Project period: 01.01.2012 until 31.12.2014 Homepage: www.sichere-ruhr.de The river Ruhr is of important local value for recreational activities as well as source water for drinking water treatment. Despite current prohibition it is already used as bathing water. During the course of the project possible risks to human health will be identified which enables an extensive risk analysis. Concepts, how a temporary use as bathing water can be realised, are developed taking different stakeholder groups into account. Insights gained by this project can be transferred to rivers used for similar purposes. Contribution of Biofilm Centre to the consortium: Occurrence of hygienically relevant bacteria in the river Ruhr and their elimination during drinking water treatment. Implementation of rapid tests and online monitoring systems in cooperation with the IWW Water Centre. Selected facultative (Aeromonas spp, Pseudomonas aeruginosa) and obligate pathogenic (Campylobacter spp., Salmonella) organisms are quantitatively determined using cultural methods. Because sampling takes place in close intervals seasonal trends and peak loads can be defined e.g. heavy rain events. Furthermore the sides chosen also take different contamination sources into account. Regarding drinking water treatment several treatment steps in an associated plant are examined for their elimination efficiencies of abovementioned organisms. The establishment of the online monitoring systems using the mbOnline-Coligard® device as well as quantitative PCR for indicator organisms is an important fundament for early warning strategies. Person in charge: Marina Horstkott, M. Sc. Email: [email protected] Phone: 0049 (0) 201-1836604
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Recognition and countermeasures to transiently non-culturable pathogens in drinking water installations
Collaborative Project, Project Partners: University of Duisburg-Essen, University of Bonn, Technical University of Hamburg-Harburg, DVGW, Technical University of Berlin, IWW Water Centre Mülheim Coordination: Prof. Dr. Hans-Curt Flemming Funded by: Federal Ministry of Education and Research Project period: 01.09.2010 until 31.08.2013 Homepage: www.biofilm-management.de
Project of Biofilm Centre within the consortium: How do hygienically relevant microorganisms enter the VBNC state, how do they become culturable again and how virulent are they then?
Copper plumbing materials can be the source of copper ions in drinking water supplies. It can be expected that P. aeruginosa and L. pneumophila may occur in the VBNC state in copper plumbing systems and thus not detectable by culture-based methods routinely employed for monitoring drinking water quality. This matter is of hygienic relevance if VBNC-cells remain cytotoxic or at least are able to regain it. Recently published results indicated that copper ions in concentrations relevant to household plumbing systems can induce a VBNC-state in P. aeruginosa (Dwidjosiswojo et al. 2011) and L. pneumophila (not published). Not relying solely on culture-dependent methods, two culture-independent techniques, fluorescent in situ hybridisation (FISH) and the LIVE/DEAD BacLight differentiation system, were applied to investigate the physiological states of the copper-treated vs. untreated cells. The transition of P. aeruginosa cells into the VBNC-state is accompanied by a loss of cytotoxicity in Chinese hamster ovary (CHO-9) cells as well as human lung epithelial (BEAS-2B) cells (cooperation with the IWW Water Centre), but P. aeruginosa can regain both culturability and cytotoxicity, when copper stress is abolished by adding a copper-chelator (Dwidjosiswojo et al. 2011). This particular trait has yet to be demonstrated for L. pneumophila. Current experiments deal with the effect of copper on established biofilms. Here the physiological states of biofilm cells including planktonic cells are investigated using both established methods FISH and LIVE/DEAD, as well as by applying new techniques to quantitatively determine ATP-content and to analyse the dehydrogenase activity with the XTT viability assay. Further important approaches to the VBNC-topic which are currently in study or in planning are the pH-dependency of the copper inhibitory effect, the investigation of gene-regulation affected by copper (microarray) and a possible resuscitation of L. pneumophila induced by Acanthamoeba.
In charge: Zenyta Dwidjosiswojo, e-mail: [email protected], phone: +49-201-183-6613
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Nanosilver particles - mechanisms of action and study of possible interaction with tissue, cells and molecules. Definition of their relevant potential for intolerance.
Collaborative Project - Project Partners: aap Biomaterials GmbH, EXcorLab GmbH, Justus Liebig Universität Gießen, rent a scientist GmbH Coordination: C. Sattig, aap Biomaterials GmbH Funded by: Federal Ministry of Education and Research Project period: 01.10.2010 until 30.09.2013 Homepage: www.nanosilver-project.info The toxic effect of silver has been known for a long time. With increasing resistance of microorganisms against antibiotics, the use of silver has gained interest in medical applications. Nanomaterials such as nanosilver (AgNPs) may have new and different characteristics in contrast to the bulk material. Until today little is known about possible side effects of nanomaterials towards humans. Substances that are categorized as well-tolerated can exhibit toxic side effects, if they are present as nanomaterial. Aim of this study is to investigate possible concerns associated with AgNPs referring to long-term effects in humans as well as possible silver resistance of pathogens.
Project of Biofilm Centre within the consortium: Antimicrobial effect of AgNPs and possible generation of microbial resistance
Aim of this study is to obtain information about the efficiency of AgNPs against clinically relevant bacteria in order to evaluate application spectra and limitations. It was found that AgNPs are less effective towards the inhibition of biofilm formation and the inactivation of established biofilms than silver ions. Attached biofilm bacteria are less sensitive towards silver (AgNP and AgNO3) than planktonic bacteria. One response of microorganisms to silver stress may be the transition into a viable-but-nonculturable- (VBNC-)state, from which microorganisms are able to resuscitate and therefore are able to regain infectivity. An indication for the VBNC-state in silver exposed Pseudomonas aeruginosa was investigated within this project. P. aeruginosa, that is not culturable anymore, still shows presence of ribosomes (FISH), intact DNA (DAPI) and the presence of Adenosintriphosphate. Furthermore microorganisms may possess, acquire or develop resistance mechanisms against silver in order to remain metabolic activity and the ability to proliferate. It is known that mechanisms responsible for heavy metal resistance also may be responsible for antibiotic resistance. Therefore the potential of silver and antibiotic resistance will be analyzed. Person in charge: Alexa Pillen, M.Sc.
Email: [email protected] Transmission Electron Micrograph of Phone: 0049 (0) 201-1836604 Silver Nanoparticles
20 nm
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Microbial ochering in technical systems
Collaborative Project - Project Partners: TU Berlin, Friedrich Schiller University Jena, HTW Dresden, Berlin Centre of Competence for Water, Berliner Wasserbetriebe, Institute for Scientific Photography, Hammann GmbH, ARCADIS Deutschland GmbH, AUCOTEAM GmbH, KSB Aktiengesellschaft, RWE Power AG, VATTENFALL Europe Mining AG Funded by: Federal Ministry of Education and Research Project period: 01.02.2011 until 31.01.2014 Homepage: www.anti-ocker.de
In water-bearing technical systems, particularly in wells or pipe networks, microbial influence may cause the formation of clogging materials. These precipitates negatively affect the per-formance of wells by blocking the screens and the pump. The aim of the project is to gain a better understanding of the process of microbial clogging and to develop new techniques to prevent or dissolve the incrustations.
Contribution of Biofilm Centre to the consortium: Integration, persistence and control of hygienically relevant bacteria in iron oxide incrustations in wells
Groundwater can become contaminated by hygienically relevant microorganisms, e. g. in consequence of heavy rainstorms, flooding of wells or hydraulic short circuits between sur-face water and groundwater. In case of an entry of hygienically relevant microorganisms into the aquifer, these microorganisms can reach drinking-water wells. If incrustations like oxides or oxyhydroxides of Fe(III) exist in the water well, the microorganisms encounter a very large and porous surface. Aim of the project is to answer the following questions
If incrustations exist in a drinking-water well, can this matrix function as a habitat? Could hygienically relevant microorganisms integrate in these precipitates? Do they persist or grow there?
If so, how can these microorganisms be eliminated most efficiently and lasting?
Analyses for hygienically relevant bacteria by culturing methods revealed coliform bacteria and Aeromonas spp. in ochre from dewatering wells of open pit mines. Survival of target organisms (E. coli, intestinal enterococci, coliform bacteria, Legionella pneumophila, Pseudomonas aeruginosa, Aeromonas spp.) spiked in ochre and well-water is tested over time in batch experiments and under flow through conditions. Batch experiments showed:
No increase of target organisms in ochre or well water
But all target organisms still culturable after 14 days
Decrease in ochre suspension less than in water show: E. coli, K. pneumoniae, E. faecalis and P. aeruginosa, but not: A. hydrophila and L. pneumophila
Different culturability of the target organisms in suspensions of different ochres over time
Person in charge: Barbara Dericks, Dipl.-Biol., M.Sc.
Email: [email protected] Phone: 0049 (0) 201-1834281
Column experiment to test survival of target organisms in ochre under flow through conditions
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Dynaklim (“Dynamic adaptation of regional planning and developmental processes to the impact of climate change in the Emscher-Lippe region (Ruhrgebiet)”
Joint project, partners: IWW Water Centre
Funded by the Federal Ministry of Education and Research
Homepage : www.dynaklim.de
Contribution of Biofilm Centre within the project:
The task of the Biofilm Centre is to perform research on the influence of temperature on hygienically relevant microorganisms in drinking water biofilms in close collaboration with the IWW Water Centre (Mülheim/Ruhr).
State of affairs
Field Experiments
Variations in temperature of about 10 °C at the sampling sites could be detected in summer. These variations were climatope-dependent and more intense in highly sealed areas (e.g. city center). For the tested drinking water no temperature effect on the detection of hygienically relevant microorganisms could be observed. Compared to the drinking water, more hygienically relevant bacteria could be detected in drinking water biofilms. Hereby, the frequency of detection was dependent on the material (EPDM> PE> stainless steel). Moreover, a temperature effect was observed for the frequency of detection of Escherichia coli and coliform bacteria (summer> autumn> winter).The total cell count slightly increased with elevating temperatures. By means of Fluorescence in situ hybridization (FISH) the target organisms could be detected at all sampling sites at least for one sampling time point. This might be an indication for bacteria being present in a VBNC state. However, this has to be further investigated.
Laboratory Experiments
All target organisms could be successfully incorporated into previous established biofilms. No effect of nutrient availability, temperature or material on culturability could be observed for Pseudomonas aeruginosa. For E. coli and coliforms an enhancing effect of nutrient addition on culturability could be observed. For temperatures above 21 °C the concentration of culturable E. coli and Klebsiella pneumonia increased while numbers of FISH-positive cells remained constant. This might be due to a transition from the VBNC state and/or multiplication. In addition to the biofilms the effluents of the annular reactors were analyzed. In 100 mL Volume of the reactor effluents the target organisms could be detected for the same time as they were detected in the biofilms indicating a contamination of the water phase by detachment of the hygienically relevant bacteria from the biofilm.
Current Experiments
To further examine if bacteria are present in a VBNC state, a DVC-FISH method was established. It could be successfully applied for drinking water samples, but needs to be optimized for the detection of bacteria in drinking water biofilms. Moreover, a PCR method will be established for the detection of E. coli in drinking water biofilms. Specificity of the oligonucleotide probe LEGPNE1 (specific for Legionella pneumophila) is investigated by application of a competitor probe (LEGCOMP2).
Person in charge: Janine Wagner, M.Sc.
e-mail: [email protected] Field exposure of stainless steel coupon holders Phone: 0049/(0)201/183-6612 (upper pipe) in an operating drinking water system
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BOSS – Biofilm Organisms Surfing Space.
Collaborative Project - Project Partners: DLR Cologne (Germany), CEPSAR (UK), University of Rome (Italy), University of Salzburg (Austria) Coordination: Dr. Petra Rettberg, DLR Cologne Funded by: Federal Ministry of Economics and Technology Project period: 01.05.2011 until 30.04.2014
The core hypothesis of the project ‘BOSS’ is the assumption that biofilm-forming organisms embedded within self-developed extracellular polymeric substances (EPS matrix) are more resistant to environmental stressors as they exist in space and on Mars compared to their EPS-free planktonic counterparts. Various test organisms such as extremophilic bacteria will be subjected to the environmental conditions as they exist in space and on Mars. This project investigates the influence of the biofilm matrix on the survival of Deinococcus geothermalis after exposure to space and Mars conditions (desiccation, UV irradiation, vacuum or simulated Martian atmosphere). Samples of water-unsaturated biofilms have been prepared using various cultivation conditions targeting highest EPS yield. Different carrier materials were tested in order to find a setup which resists the detrimental effects of space vacuum and irradiation.
The effects of long-term desiccation on the viability of planktonic and biofilm cells were studied using cultural and fluorescence-microscopic techniques. In order to be able to identify possible EPS-mediated resistance mechanisms, EPS from biofilms grown on membrane filters were isolated using various techniques and characterised in terms of their composition (proteins, total carbohydrates, uronic acids, extracellular DNA) using photometric assays. First results which already support the project hypothesis have been presented at the 12th annual meeting of the European Astrobiology Network Association (EANA 2012) in Stockholm, and at the international conference Biofilms V in Paris.
Live/Dead-stained biofilm of D. geothermalis desiccated for 90 days, indicating well preserved membrane integrity (1,000X).
In collaboration with the working group of Prof. Christian Mayer at the University of Duisburg-Essen, the effects of desiccation on the mechanical properties of biofilms are investigated.
Person in charge: Jan Frösler, M.Sc.
Email: [email protected] Phone: 0049 (0) 201-1836613
Technical assistence: Anika Marko Email: [email protected] Phone: 0049 (0) 201-1836618
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Proteins, polysaccharides and nucleic acids as components of the extracellular polymeric substances of drinking-water biofilms Financial support: Max Buchner Research Foundation Project period: 01.02.2008 until 06.11.2012 (The Project is finished) Coordination: Prof. Dr. Hans-Curt Flemming
Project description
Drinking water distribution systems as well as domestic plumbing systems are colonized by microbial biofilms. Under unfavourable conditions they may act as reservoirs for hygienically relevant microorganisms, posing a potential threat to human health. The aim of this study was the investigation of the formation, composition and function of drinking-water biofilms and their extracellular polymeric substances (EPS). The biofilms were grown on a synthetic elastomeric material, exposed to drinking water in different drinking water distribution systems as well as public plumbing systems made of copper. Characterization of biofilms and their EPS was carried out by microbiological, molecular biological and biochemical methods, which were adapted and optimized to meet the difficulty provided by the low amounts of biomass usually found in drinking water systems. The results of this study demonstrated a strong variability among drinking-water biofilms in terms of microbial populations and EPS composition in response to variations of conditions in different distribution systems and in particular in copper plumbing systems. Furthermore, this study demonstrated the dynamics of EPS components in the course of biofilm formation, indicating continuous changes to the EPS matrix induced by the constituent organisms. Drinking-water biofilms were shown to be another type of biofilms, in which proteins represent the main EPS component, followed by polysaccharides and DNA. EPS proteins in drinking-water biofilms exhibited metabolic, transport and regulatory functions. Person in charge: Witold Michalowski, Dr. rer. nat.
Successful completion of the PhD project Two-dimensional gel of EPS proteins of a 14 d-old drinking-water biofilm Email: [email protected] Phone: 0049 (0) 201-1836605
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Freshwater plankton as a habitat for hygienically relevant microorganisms
Funded by: DFG (German Research Foundation)
Project period: 01/2009 until 12/2012
Collaboration: NRF-funded project: “Plankton as a vector for hygienically relevant bacteria in eutrophic waters”; Prof. Dr. T. E. Cloete, University of Stellenbosch, South Africa
In aqueous environments bacteria can occur planktonically in the water phase, or associated in biofilms attached to solid surfaces or other phase boundaries. During high proliferation periods in summer, plankton organisms in surface waters provide external surfaces which can be colonized by biofilms. Zooplankton surface can add up to an overall area of more than 3.000 km2 in a lake. Possible associations of potentially pathogenic bacteria with phyto- and zooplankton were observed in a field study in a freshwater environment (Lake Baldeney, Essen, Germany) and in laboratory experiments. Hygienically relevant microorganisms considered were, bacteria with faecal origin (Escherichia coli, coliforms, intestinal enterococci, Clostridium perfringens), an obligate human pathogen of faecal origin (Campylobacter spp.), and environmental opportunistic bacteria (some coliforms, Pseudomonas aeruginosa, Aeromonas spp., Legionella spp.). For all of the investigated hygienically relevant bacteria an association with freshwater plankton could clearly be demonstrated, except for Campylobacter spp. which was only found in water and Legionella spp. which was not detected by culture. The bacterial abundance in all samples was found to be higher with plankton than compared to the free water. With the culture-independent methods, fluorescence in situ hybridization (FISH) and quantitative polymerase chain reaction (qPCR), the organisms P. aeruginosa and Legionella spp. were found in significant higher concentrations in water and plankton than with cultural methods. This observation indicates that both may occur in a viable but nonculturable (VBNC) state. In laboratory experiments hygienically relevant bacteria were co-cultivated with Daphnia magna in microcosms. To study cladoceran-bacteria associations in detail, it was discriminated between carapace-associated and ingested bacteria. The organisms P. aeruginosa and A. hydrophila were found more frequently located on the carapace of D. magna, whereas most of E. faecalis was located in the gut. FISH analysis indicated the possibility of VBNC cells for these organisms in association with D. magna. The field study as well as laboratory microcosms indicate that there are associations and accumulations of pathogenic bacteria. Plankton can act as a reservoir and a vector for potentially pathogens and may spatially enhance bacterial concentrations up to infectious doses. In case of VBNC bacteria, it is possible that the VBNC cells resuscitate and regain their virulence. Plankton-pathogen associations are of relevance considering human health in drinking water production and recreational use of the surface water.
Person in charge: Miriam Tewes, MSc
Email: [email protected]
Phone: 0049 (0) 201/183-6612
SEM micrograph D. magna
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Advanced Technologies for Water Resource Management (ATWARM) Marie-Curie project
Collaborative Project - Project Partners: QUESTOR centre (UK), ATWARM, University of Duisburg-Essen, Queen’s University Belfast (UK), Cranfield University(UK), IWW Water Centre, Industrial partners: NI Water (UK), T.E. Laboratories Ltd (Ireland). Coordinator: Prof. Mike Larkin, Queens University, Belfast Homepage: http://www.atwarm.com
Contribution of Biofilm Centre and Institute of Analytical Chemistry to the consortium within the project: Priority substances in activated sludge: Incidence, accumulation, source tracking emitter identification & prevention strategies
This project is aimed at finding innovative applications of microbial wastewater biofilms. Due to their ability to interact with many organic and inorganic pollutants, waste water microbial biofilms potentially represent a useful tool in order to trace pollution processes in waste water piping systems and to prevent pollution events. Biofilms can absorb certain pollutants and after a certain period of time release them back into the environment- The main part of the project is to study the sorption kinetics that preside the processes of absorption and desorption of pollutants from the biofilm matrix, firstly using artificial biofilm- gels and then, of course, testing the real biofilms present in the waste water system.
The main pollutant taken as a reference compound in this research project is phenanthrene, one of the most studied PAH compounds (Poly Aromatic Hydrocarbons). During the first part of the project the aim will be to study the absorption and desorption processes of phenanthrene in a gel matrix, used as a biofilm model. Fluorescence spectrometry will be used as reference technique in order to perform this investigation.
Results Quartz cuvettes have been used to perform a front phase fluorescence analysis of the sorption properties of agar gel matrices against phenanthrene water solutions at 1.5ppm concentration, 1:1 volume ratio. Mixing and static conditions have been tested and slow diffusion processes have been revealed. Mixing condition in water phase has been performed with a speed of 900rpm by magnetic stirring and the results show that in both processes, absorption and desorption, the mass transfer values are higher than the static condition of the experiments. Maximum value of 6.0x10-8 g/m²s has been reached in static conditions and a maximum value of 7.0x10-5 g/m²s in mixing conditions.
Person in charge: Giacomo Bertini, M.Sc.
[email protected] 0049-15208371712
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4.0x10-8
6.0x10-8
8.0x10-8
1.0x10-7
Mass flow
Decay 1
F (g
/m²*
s)
days
Absorption agar 2%
Static cuvette
3.5mL quarts cuvettes have been used to study the absorption of phenanthrene by a gel matrix made of agar (2%) . The results reveal a slow diffusion process reaching the equilibrium with a distribution coefficient equal to 1.
17
Membrane biofouling: hydraulic resistance of biofilms
Funded by: Wetsus, centre of excellence for sustainable water technology Project period: 01.04.2009 until 30.06.2013 Supervisors: Prof. Dr. Hans-Curt Flemming and Dr. Hans Vrouwenvelder Homepage: http://www.wetsus.nl/research/research-themes/biofouling Membrane filtration is one of the most common procedures for effective water production and can be used in many different processes such as drinking water production and waste water treatment. These filtration systems suffer from performance losses due to fouling which may have different sources like scaling, organic fouling, particle fouling or biofouling. Biofouling is a big issue in membrane filtration systems because it is caused by bacterial cells which, once attached to the membrane, form biofilms.
Biofilms consists of bacterial cells embedded in a highly hydrated matrix of extracellular polymeric substances (EPS).The biofilm permeability is influenced by the operating conditions of the system, such as cross and permeate flow velocity and nutrient availability.
The aim of this study is to investigate the contributing factor for high and low hydraulic biofilm resistance and its impact on different filtration systems.
A lab scale crossflow filtration system consisting of four flow cells is operated under various conditions: different cross and permeate flow velocities as well as feed water supplemented with different substrate types and concentrations. The obtained biofilms are analyzed for total cell number, protein and polysaccharide concentration, total organic carbon and thickness.
These studies were performed with microfiltration membranes (0.05 µm pore size) to rule out the effect of concentration polarization and permit the study of pure biofilm resistance. It was demonstrated that no fouling occurred inside the pores of the membrane. The biofilm resistance (6x1012 m-1) found in the filtration system was high compared to the resistance of the employed MF membrane (5x1011 m-1) but much lower than the resistance of an NF 2x1013 m-1) or RO membrane (9x1013 m-1). This indicates that fouling of NF/RO systems can not be ascribed exclusively to biofilm formation.
Furthermore, high permeate flux (100 Lm-2h-1), high crossflow velocity (0.2 m-s) and the presence of a feed spacer increase biofouling and biofilm resistance. Nutrient availability in the feed water presents the most important reason for increased biofouling and rules out the influence of all other operational parameters (Dreszer et al., 2013).
Person in charge: Claudia Dreszer, M.Sc.
[email protected] Scanning electron micrograph of a microfiltration Phone: 0031 (0) 582843183 membrane covered with biofilm
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Iron- and sulfur- oxidizing microorganisms in the Lusatian- and Central German lignite mining districts
Originaltitel: Eisen- und Schwefel-oxidierende Mikroorganismen im Lausitzer und Mitteldeutschen Braunkohlerevier
Collaborative Project - Project Partners: Hochschule Lausitz (FH) University of Applied Science; Universität Duisburg-Essen (UDE), GMB GmbH; MIBRAG mbH; Vattenfall Europe Mining AG Persons in charge: Bianca Florian, MSc; Sören Bellenberg, MSc; Prof. Dr. W. Sand Project Funding: GMB mbH, MIBRAG AG, Vattenfall Europe Mining AG Project period: 01.01.2008 until 31.12.2011 Bioleaching, the dissolution of metal sulfides such as pyrite and chalcopyrite, occurs by
microbial activity. Acid mine drainage (AMD) is therefore a natural process. Wherever metal
sulfides are exposed to oxygen and water, as it occurs when iron sulfide enclosures in coal
are made accessible for leaching bacteria during mining. It has been found that attachment
of leaching bacteria to the mineral surfaces enhances the process of metal sulfide dissolution
and AMD generation. Therefore, in this study we quantify and visualize the initial colonization
and biofilm formation of leaching microorganisms on sulfide minerals. Biofilms are visualized
by epi-fluorescence microscopy (EFM) using DAPI, SytoTM 9, FISH, lectin- and calcofluor-
staining. Additionally, atomic force microscopy (AFM) is used for the investigation on cell
morphology, spatial arrangement of cells on pyrite and mineral surface topography. The
ensuing work is concerned with inhibition measures in order to reduce AMD/ARD as much as
possible. Leaching-inhibition with several detergents was demonstrated in column
experiments with material from lignite mining areas, where AMD/ARD takes place.
Microcalorimetry and chemical monitoring of metal sulfide dissolution products was used to
determine metabolic activity and its inhibition. Detergents efficiently inhibited leaching and
cell lysis resulted in reduced cell numbers in effluents and on mineral surfaces. Application of
detergents to prevent leaching during mining activities is considered in the future. Therefore
after the project, results shall be used for assessment of a field scale application of
detergents. Dr. Bianca Florian (UDE) and Dr. Hans-Michael Siebert (FH Lausitz) successfully
defended their project-related work and obtained their PhD-degrees in May and August 2012.
Persons in charge:
+
2.5 µm
Bianca Florian, MSc [email protected] Tel: 0049 (0) 201 183 7085
Biofilm on a pyrite surface visualized with combined atomic force and epifluorescence microscopy
Column reactor Sören Bellenberg, MSc
[email protected] Tel: 0049 (0) 201 183 7084
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Attachment to and dissolution of metal sulfides by moderately thermophilic archaeon Ferroplasma
Funded by: China Scholarship Council/Universität Duisburg-Essen
Running time: 2011-2014
The attachment of microorganisms to mineral surfaces is postulated to play a significant role in the mineral dissolution process. Extracellular polymeric substances (EPS) mediate the connection between cells and the mineral sulfide and play a crucial role in interfacial interactions. Ferroplasma spp are extremely acidophilic iron-oxidizing archaea and always present in acid mine drainage (AMD) and bioleaching systems, and are thought to paly important role in global iron and sulfur cycling.
Contribution of Biofilm Centre within the project: Investigation of the process of attachment to different mineral surfaces, extraction and analysis of EPS involved in attachment and biofilm formation on sulfide minerals. We intend to provide new insights about the attachment of archaea to metal sulfides in order to deepen the understanding of the interfacial mechanism of microbial leaching and its key factors.
Work in Progress: (1) Investigations of biofilm formation of F. acidiphilum by advanced microscopical techniques during the bioleaching process are continued. By now AFM combined EFM studies showed that: F. acidiphilium biofilms increased with incubation time and a monolayered biofilm was formed on pyrite surfaces. Pits were found and large pyrite surfaces were not colonized by cells after up to two months of incubation. Cells showed preference to attach to the cracks / defects of the pyrite. (2) Use of lectin-binding analysis allowed us to provide new details on the composition of extracellular polysaccharides produced by the organism, suggesting that F. acidiphilum may contain mannose, N-acetylgalactosamine and L-fucose in the surface polysaccharides. (3) Contribution of F. acidiphilum to the mineral dissolution was compared with the activities of the well-recognized leaching bacteria such as Acidithiobacillus ferrooxidans and Leptospirillum ferriphilum, demonstrating relatively low capacity of pyrite dissolution. (4) Updated results of biochemical analysis showed that colloidal EPS mainly contained sugars however capsular EPS contained sugars and proteins. Further investigations are needed to find out the links between functional and biochemical properties of archaeal biofilms involved in bioleaching of pyrite. Person in charge: Ruiyong Zhang (M.Sc.)
Archaeal biofilms on pyrite visualized by atomic force microscopy (left) and epifluorescence microscopy (nucleic acids staining, middle and EPS staining right)
0049 (0) 201-183-7076
20
Urban mining – Recovery of metals from lignite ashes
Original title: r3 - Strategische Metalle , Verbundvorhaben: Kraftwerksasche - Chemisch-biotechnologische Gewinnung von Wertstoffen aus BraunkohIenkraftwerksasche. TP 3 - Mikrobielle Laugung mit thermophilen Schwefeloxidanten und Nitrifikanten Joint project - Project Partners: − G.E.O.S. Ingenieurgesellschaft mbH; Prof. Dr. A. Schippers, BGR (Bundesanstalt für
Geowissenschaften und Rohstoffe); Prof. Dr. W. Sand, Universität Duisburg – Essen; Prof. M. Bertau und Prof. G. Heide TU Bergakademie Freiberg; Prof. H. Wotruba RWTH Aachen; Prof. Dr.-Ing. Horst-Michael Ludwig, Bauhaus-Universität Weimar; Nickelhütte Aue GmbH; Loser Chemie GmbH; Vattenfall Europe Mining AG; Sunicon GmbH Persons in charge: Sören Bellenberg, MSc; Dr. Tilman Gehrke, Prof. Dr. W. Sand Project Funding: BMBF Project period: 01.011.2012 until 31.10.2015
Bioleaching is well suited for future application in recycling and recovery of metals due to heavy metal tolerance and general extremophilic nature of acidophilic microorganisms. Raw material supply is a mayor issue for ensuring success of the European industry, economy and society. Scarcity of these resources demands economical use and causes high market prices. Critical metal resources are essential for almost all high technology electronic products. Lignite fired power plants are a backbone of the energy production in Germany. Tremendous amounts of ashes were landfilled in the past. Due to their metal contents, lignite ashes are now considered as a potential novel resource. The amounts and metal contents will be encompassed and samples will be processed and selected for biological treatment steps. Screening for heavy metal resistant microorganisms suitable for metal solubilization from ashes is one aim of this project. Extreme acidophilic, chemolithoautotrophic, sulfur and iron oxidizing bacteria and archaea will be used. The potential of nitrifying and organic acid producing microorganisms for this purpose will also be investigated. Metal solubilization from lignite ashes and concentrates shall be achieved in order to develop a geobiotechnological metal winning process.
Persons in charge:
Sören Bellenberg, MSc [email protected]
Tel: +49 201 183 7085
Dr. Tilman Gehrke [email protected]
Tel: +49 201 183 7081
21
Microcalorimetry – a Measuring Technique for the Detection of Biomining
Funded by: Federal Institute for Geosciences and Natural Resources Project period: 01.10.2010 until 31.12.2014 Collaborative Project - Project Partners: Federal Institute for Geosciences and Natural Resources & Universität Duisburg-Essen, Biofilm Centre, Aquatic Biotechnology The demand (price) of metals especially copper is increasing since many years. The mining techniques for the recovery of metals are cost intensive and environmentally harmful. High grade ore deposits are running off and low grade ores as well as recycling of metals have to be considered as resources. Compared to pyrometallurgical treatment, biomining is an environmentally friendly, cheap and simple mining technique. Microorganisms are solubilizing minerals from their ores. Since more than 40 years biomining is economical used for the recovery of metals from low grade ores to win metals like copper, nickel and uranium. Contribution of Biofilm Centre to the consortium: Development of a microcalorimetric measuring technique for biomining Aim of this study is the development of a measuring technique for the quantification of biomining of three different copper sulfides (Chalcopyrite, Chalcocite, Covellite). Microcalorimetry is a non-invasive, non-destructive analytical technique, which provides information about the metabolic activity of microorganisms. In combination with analytical techniques, which can track the chemical products produced during the bioleaching process, the bioleaching efficiency may be quantified and predicted. The copper sulfides will be leached with microorganisms in different temperature ranges. Microcalorimetric measurements will be performed to investigate the microbial activity and Atomabsorptionspectroscopy, High-Pressure Liquid Chromatography, Ion Chromatography and UV/VIS Photometry will be performed to analyze the reaction products copper, sulfur, sulfate & thiosulfate and iron. Person in charge: Beate Krok For efficient bioleaching, the attachment of microorga- [email protected] nisms to the ore is required. The attachment is mediated 0049 (0) 201 183 7089 through the extracellular polymeric substances (EPS)
layer produced by microorganisms. The EPS plays also a crucial role as reaction space for the bioleaching process. To investigate the interactions of microorganisms with mineral grains, the combination of Atomic Force with Epifluorescence Microscopy will be used. Furthermore, the microcalorimetric measuring techniques will be verified by samples from copper deposits. The experiments show that a sufficient adaptation of microorganisms to copper sulfides, especially chalcopyrite is needed before the conducting of bioleaching experiments. In general themophilc microorga- nisms show a higher potential for the extraction of copper from copper sulfide minerals. TAM Microcalorimeter
22
Nutrition optimization of thermophilic and halophilic phototrophic microorganisms
Funded by: YARA GmbH & Co. KG
Project period: 01.07.2012 until 31.07.2013
Project description
The origins of cyanobacteria date back almost to the beginning of life on earth. The threat posed by greenhouse gases and global warming to both human and nature is a worldwide issue and is deemed as one of the major challenges that must be solved in the coming decades. Further aspects are the actual situation about world nutrition and the generation of alternative energy sources. For various photo bioreactors operating at different places in the world there is a need of suitably adapted microorganisms.
Particularly, this project is focused in the search and characterization of cyanobacteria suitable for a pilot plant which is operated with seawater and located in desertic environments. Consequently, these microorganisms shall be able to grow in sea water and high temperature environments.
In the pilot plant open pond reactors will be used. One of the major advantages of open pond systems is that they are easier to construct and operate than most closed systems. The major limitations in these systems include poor light utilization by the cells, evaporative losses and requirement of large areas of land. Furthermore, contamination by predators and other fast growing heterotrophs have restricted the commercial production of algae in open culture systems to only those organisms that can grow under extreme conditions.
Additionally, the determination of the optimal micro- and macro nutrients as well as an optimization of the addition of fertilizers in relation to the increasing biomass will be investigated.
Macro and micro nutrients in complex matrices, from batch cultures in different growth states, are determined by Flame or Graphite Furnace AAS. Based on the previously obtained results an optimized nutrition will be offered.
Person in charge: Inga Vanessa Kirstein, B.Sc.
Photomicrographs of the culture strain Iceland clone 2e, at 45°C and in IOBG-11 medium (4% salt concentration), scale bar = 5µm (a) DAPI stained fluorescing trichomes (b) autofluorescence of the cyanobacteria (c) mixture of DAPI and auto- fluorescence.
Email: [email protected]
Phone: 0049 (0) 201-183 7085
23
BIOCOR Initial Training Network (ITN) on biocorrosion
Research progress in 2012 within the Individual Project (IP) No. 4 (UDE, Biofilm Centre): The role of EPS in biocorrosion initiation or inhibition.
Joint project, partners: The project involves 12 research teams and 4 associated partners from 9 different EU countries, from both academic and industrial sectors
Funded by: EU Seventh Framework Program "People"
Project period: 1.09.2009 – 1.09.2013
Homepage: www.biocor.eu
Biofilm growth mode of Microorganisms, in a majority of their habitats causes a number of serious industrial problems by altering the interfacial chemistry between the colonized material and the bulk fluid. Corrosion of low carbon steel, material widely used in the oil transport and processing systems can be strongly accelerated by microorganisms from various physiological groups. Sulfate reducing prokaryotes (SRP) are considered as probably the most aggressive ones. Although the impact of their biofilms on the metal surface has been extensively studied, the role of extracellular polymeric substances (EPS) in metal–microbial interactions still remains unclear. Hence the main objective of IP 4 was focused on the investigation of EPS role in the MIC processes. In order to bring deeper insights into the functionality of EPS a comprehensive approach, performed in collaboration with other BIOCOR ITN partners (Universidade Nova de Lisboa (UNL), Portugal and Université Catholique de Louvain (UCL), Belgium) was applied. Different microscopic techniques (Atomic Force Microscopy (AFM), Confocal Laser Scanning Microscopy (CLSM), Transmission Electron Microscopy (TEM)), electrochemical analyses, as well as surface studies (X–ray Photoelectron Spectroscopy (XPS), Time of Flight Secondary Ion Mass Spectrometry (ToF–SIMS) allowed us to understand the way EPS contributes to the biofilm formation and its interactions with the metal surface. Monitoring of Ecorr showed that the early stages of the biofilm formation were critical for the development of corrosion processes. The main consequence of microbial activity on the carbon steel surface was the development of the localized corrosion in the form of pitting, as confirmed by AFM analyses. CLSM monitoring of the biofilm formation suggested the leading role of proteins in this process. Proteomic studies of EPS allowed detecting above 100 proteins in the extracellular matrix. Among them heme containing molecules (possibly cytochromes) were found, suggesting possible involvement of EPS in microbe – metal interactions. The results of CLSM investigations were confirmed by XPS characterization of extracellular matrix on the different surfaces (glass and polystyrene as the model hydrophilic and hydrophobic surfaces, and the carbon steel for the field investigations). Extracellular molecules attached to the different surfaces generally included 70 to 80 % of proteins, 10 to 20% of hydrocarbons (lipids) and up to 20% of sugars or eventual contaminants. Combination of the methods mentioned gives better overview on the complex problem of biofilm formation by corrosion relevant prokaryotes on the industrial material.
Persons in charge:
(a) (b)
M.Sc. Agata J. Wikieł
Tel: 0049 (0) 201 183 7083
M. Sc. Iaryna Datsenko
Tel: 0049 (0) 201 183 7083
XPS characterization of EPS produced by D. alaskensis AL 1 (a) and the pitting corrosion of carbon steel caused by the
bacterial biofilm (b)
24
Biogenic sulfuric acid corrosion of different materials
Project Partner: Fraunhofer Institute for Environmental, Safety and Energy Technology UMSICHT in Oberhausen
Biogenic sulfuric acid corrosion is a chemical attack to surfaces of different materials such as concrete, iron and polymers. It is caused by sulfuric acid producing bacteria from the genus Thiobacillus. It mainly occurres in waste water systems, where sulfur compounds are degraded by microorganisms. Gaseous sulfur compounds are released and accumulate in the head space. Chemical oxidation of H2S to elemental sulfur and the following biological oxidation via thiosulfate and other polythionates cause a decrease of pH (<7). The reduced sulfur compounds are oxidized to sulfuric acid, resulting in suitable conditions for growths of Thiobacilli (T. neapolitanus, T. intermedia) which further decrease the pH. Below pH 5.5 A. thiooxidans colonize the surface. Between pH 2.0-3.0 this organism will find optimal growth conditions. Result is a successive colonization of surfaces by different Thiobacilli. Sulfuric acid is produced as a metabolite of these organisms causing an attack on different materials.
Aim of this study is the optimization and further development of different materials against biogenic sulfuric acid corrosion
Contribution of Biofilm Centre to the consortium: Microbiological and analytical investigation of materials which were stored in the pilot plant at Fraunhofer Institute
In cooperation with Fraunhofer Institute for Environmental, Safety and Energy Technology UMSICHT in Oberhausen a pilot plant was developed and the effect of biogenic sulfuric acid corrosion on different materials is tested. The pilot plant is located at the Fraunhofer Institute, microbiological analyses are performed in the lab of the Aquatic Biotechnology, Universität Duisburg-Essen. All samples are investigated using analytical as well as microbiological techniques. Microbiological analysis include different organisms like acidophilic and neutrophilic iron- and sulfur oxidizers, sulfate reducing bacteria, nitrite oxidizers, fungi, chemoorganotrophic bacteria and more.
Person in charge: Nanni Noël
Nach: Bock, E.; Sand, W.; Pohl, A.: Bedeutung der Mikroorganismen bei der Korrosion von Abwasserkanälen, TIS Tiefbau- Ingenieurbau- Straßenwesen, Sonderdruck zum 4. Statusseminar “Bauforschung- Technik“, 1983, S. 47-49
[email protected] phone: 0201-1837085
25
Bio-derived corrosion protection for metallic materials by analogues of microbial exopolymeric substances from renewable resources
AIF joint project together with: DFI – DECHEMA Research Institute Funded by: Programme for Industrial Collective Research (IGF) of the German Federal Ministry of Economy and Technology under the auspices of AIF, Project ZN04204-10 Period: 01.02.2011 – 30.06.2013
Microbial biofilms and their surrounding extracellular polymeric substances (EPS) can both proliferate and inhibit corrosion. The first is denoted as microbiologically influenced corrosion (MIC), the latter as microbiologically influenced corrosion inhibition (MICI). Both effects are influenced by the interactions between the EPS matrix and the constructional material itself, with the EPS’ chemical composition determining its detrimental or beneficial nature. Comparable to classical interfacial corrosion inhibitors, specific functional groups of the EPS are crucial for EPS-surface interactions and thus facilitation of cell adhesion. This project aims to investigate EPS-analogue substances to protect metals against (a)biotic corrosion and formation of detrimental biofilms on their surface. The blocking of electrochemically “active sites“ should suffice to prevent bacterial chemotaxis to (iron) ions and thus to reduce attachment of detrimental microbes to these sites.
Contribution of the Biofilm Centre: Extraction and analysis of microbial EPS, MIC simulation and visualization of the metal-coating-bacteria interfaces
The Aquatic Biotechnology working group is constantly growing biofilms of detrimental and beneficial microorganisms to extract and analyse their EPS. Our MIC-simulations with sulphate-reducing bacteria showed a reduction in corrosion rates of mild steel coated with analogues substances by up to 99% compared to uncoated references. A potential impact of the coatings on pitting corrosion of highly alloyed steel by Mn-oxidising bacteria is currently under investigation. Biofilm formation of both groups of organisms was only slightly reduced, but the coatings effectively prevented contact of the microorganisms to the material surface. The biodegradability of the substances by pure cultures of SRB as well as enrichment cultures from soil (“worst case”) in short-term was negligible, but their longer-term stability still has to be assessed. Currently, the influence of surfaces, bacteria and coatings on each other is studied using state-of-the art high-resolution visualisation techniques such as combined epifluorescence with both Kelvin probe force microscopy (EFM-KPFM) as well as a novel electrochemical AFM-cell.
Persons in charge:
Andrzej Kuklinski
[email protected] 0049 (0) 201-183-7082
Christian Thyssen
christian.thyssen@uni-due 0049 (0) 201-183-7075
Visualisation of D. vulgaris on mild steel
A, fluorescence image of DNA-stained cells B, topographic AFM image of the same area C, further magnification of white squared area
26
Physiological and Molecular processes involved in Biofilm formation in Bioleaching bacteria
Persons in charge: Dr. Mario Vera, MSc. Sören Bellenberg, MSc. Claudia Janosch, MSc. Nanni Noel. Prof. D Physiological and Molecular processes involved in Biofilm formation in Bioleaching bacteria Persons in charge: Dr. Mario Vera, MSc. Sören Bellenberg, MSc. Claudia Janosch, MSc. Nanni Noel. Prof. Dr. Wolfgang Sand Collaborative Project - Project Partners: Dr. Nicolas Guiliani. Laboratorio de Comunicacion Bacteriana. Universidad de Chile. Dr. Ansgar Poetsch, Plant Biochemistry, Ruhr Universität Bochum Funded by: Universität Duisburg-Essen. Programm zur Förderung des wissenschaftli-chen Nachwuchses. DAAD „Alechile“ program 2012-2013. Bacteria are able to communicate by producing and secreting a diverse set of small mol-ecules called autoinducers in a process called “Quorum sensing” (QS) regulating several population density-dependent cellular behaviors, such as virulence, conjugation, transfor-mation, and biofilm formation. Our research has shown that biofilm formation processes can be influenced by external addition of Quorum Sensing molecules. Our Methods include microscopy and molecular biology and bioinformatic studies: By using AFM, EFM and CLSM, we are imaging different polysaccharide moieties of the EPS by using fluorescently labeled lectins, following the dynamics of biofilm formation processes and the transition from planktonic to a biofilm lifestyle. We are using the current genome sequences from At. ferrooxidans, At. caldus, At. ferrivorans and At. thiooxidans, searching for genes potentially encoding EPS related proteins and trying to predict their roles on EPS formation and Biofilm formation. This data is currently being complemented through high throughput proteomic studies of At. ferrooxidans and At. caldus. In 2012, a search for new autoinducers mainly produced by biofilms of Leptospirillum, strains has started. These molecules possess inhibitory effects again several iron oxidizing acidophiles and remain to be characterized. Also the influence of the external addition of these as well as known QS molecules is used to understand the dynamics of the bacterial communication processes in leaching environments. Interestingly, some previous results suggest a strong cross-communication among species, some of them with negative effects in terms of leaching efficiencies. All these aspects have not been considered in bioleaching operations and could help in the future to improve the performance of bioleaching consortia, were positive interactions are required to enhance metal dissolution and negative ones might be suitable to inhibit acid mine drainage.
Dr. Mario Vera [email protected]
Tel: +49 201 183 7083
Sören Bellenberg [email protected]
Tel: +49 201 183 7084
Claudia Janosch [email protected]
Tel: +49 201 183 7085
Nanni Noel [email protected]
Tel: +49 201 183 7085
27
HotZyme - Systematic screening for novel hydrolases from hot environments
Funded by: 7th framework program for research and technological development (FP7) of the European Union Project period: 01.01.2012-31.12.2014 Collaborative Project - Project Partners: University of Copenhagen, MicroDish BV, Montana State University, Stiftelsen Norges Geotekniske Institut, Winogradsky Institute of Microbiology, Novozymes A/S, Wageningen University, Department ATV, University of Exeter, Sigma-Aldrich Production GmbH, Consiglio Nazionale Delle Ricerche, National Technical University of Athens, European Centre for Research & Financing Homepage: http://hotzyme.com/ HotZyme is a large-scale integrating project (EU 7th Framework) on the systematic screening for novel hydrolases from hot environments coordinated by Prof. Xu Peng (University of Copenhagen, Denmark). It recognizes that there is a strong need for new thermostable hydrolases with appropriate performance and/or novel functionalities that could provide huge savings in time, money and energy for industrial processes. HotZyme aims to identify such enzymes from hot terrestrial environments using metagenomic screening methods. New bioinformatics tools will be developed to facilitate function prediction of genes from metagenomes that show low or no sequence homology to enzymes of known function. A range of high-throughput screening technologies will be employed to identify novel hydrolases. Contribution of Biofilm Centre to the consortium: Biochemical characterization of novel enzymes Aim of this study is to understand the mechanism of action, substrate specificity and feasibility of the use of selected hydrolase enzymes for relevant bio-processes. Currently, three different thermophilic esterases are investigated: two phospotriesterases with promiscuous lactonase activity from Sulfolubus and a still uncharacterized esterase from Vulcanisaeta mountnovskia. The enzyme characterizations include the determination of stability and activity in response to temperature, organic solvent, pH and a detailed kinetic analysis. These enzymes have great potential for detoxification of chemical warfare agents and pesticides. Furthermore, lactonases are also known as quorum quenchers.
Person in charge: Dr. Verena Kallnik and Dr. Theresa Kouril
[email protected] Phone: 0201-1837068
[email protected] Phone : 0201-1837065
Environmental sampling
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Stress response on Biofilms of the thermoacidophilic Archaeon Sulfolobus acidocaldarius
Funded by: Mercator Research Center Ruhr (MERCUR) Project period: 01.03.2012 - 28.02.2015 Collaborative Project - Project Partners: Prof. Dr. Andreas Schmid, Dr. Katja Bühler, Laboratory of Chemical Biotechnology, TU Dortmund Prof. Dr. Hans-Curt Flemming, Aquatic Microbiology, University of Duisburg-Essen Dr. Jost Wingender, Aquatic Microbiology, University of Duisburg-Essen
The goal of this project is to understand the response of bacterial and archaeal biofilms
towards process related conditions relevant for the biotechnical production of chemical
materials. Biofilms as new designer biocatalysts offer the advantages of enhanced tolerance
to environmental stress factors and long-term stability, allowing for the conversion of
biologically challenging compounds and continuous processes. Two model organisms are
investigated: the mesophilic bacterium Pseudomonas sp. strain VLB120 (TU Dortmund) and
the archaeon Sulfolobus acidocaldarius (University of Duisburg-Essen). Main focus is on the
influence of solvents involved in biotransformation processes, which are most often toxic to
the biocatalyst, impairing process performance especially of planktonic cultures. When
exposed to such solvents, the intrinsic response of the cells in the biofilms is multifaceted.
Upon exposure to environmental stress, bacteria are known to enter into a viable but non-
culturable (VBNC) state. An important aspect will be to investigate the influence of solvent
stress on the vitality and physiological activity of biofilms and the possible induction of the
VBNC state of the biofilm cells. The objective is to identify and optimize the conditions for the
use of biofilms in solvent-dependent biotechnical applications.
To analyze solvent exposed biofilms, we focus on non-invasive, time-resolved and on-line
methodologies based on confocal laser scanning microscopy and flow cell cultivation
systems. In addition, biochemical assays (e.g. Lowry assay, phenol-sulfuric acid method) for
the analysis of whole biofilms and their extracellular polymeric substances (EPS) as
important structural and functional components of the biofilm matrix are widely applied in this
study. In order to investigate the influence of solvents on cellular surface structures, the AFM
facilities of the “Aquatic Biotechnology” group will be involved. Thus, this is the first project
with participation of the entire Biofilm Centre.
Person in charge: Jens Benninghoff (M. Sc.)
Email: [email protected] Phone: 0049 (0) 201 183 7067
Conceptual theme of the project.
29
ExpresSYS - A Platform of Novel Microbial Expression Systems for Industrially Relevant Genes
Funded by: Federal Ministry of Education and Research (BMBF) Project period: 01.03.2010 until 28.02.2013 Collaborative Project - Project Partners: TU München, Heinrich-Heine Universität Düsseldorf, MPI Marburg, Universität des Saarlandes, Universität Hamburg, TU Hamburg-Harburg, BRAIN AG, Evonik-Degussa, Henkel KGaA, Süd-Chemie, Wacker Chemie ExpresSYS is a collaborative project (coordinated by Wolfgang Liebl). Homepage: http://genomik-transfer.de/index.php?section=verbund&fkz=0315586 Contribution of Biofilm Centre to the consortium: A novel archaeal expression host: the thermoacidophilic Crenarchaeon Sulfolobus acidocaldarius Many archaeal species thrive in extreme habitats requiring effective adaptation and specialization strategies. Their proteins, so called “extremozymes”, are active under harsh and unreal conditions, which make them very interesting for biotechnological applications. Unfortunately, the functional expression of many archaeal (hyper)thermophilic proteins in mesophilic expression hosts or even thermophilic bacterial hosts is limited. The missing archaeal post-translational machinery, as well as the misfolding of proteins at low temperature is supposed to be a major reason. Therefore, in current metagenomic approaches only a fraction of the tremendous diversity can be accessed due to the pre-selection introduced by the choice of common bacterial expression systems. Thus, there is an urgent need for the establishment of alternative expression hosts and including an archaeal expression host is an important contribution to unravel and to exploit the biodiversity available in extreme habitats. Sulfolobus acidocaldarius is a well characterized thermoacidophilic, obligate aerobic Crenarchaeon, which grows optimally at 78°C and pH 2-3. Most important, the organism is genetically tractable and a vector system for protein expression has been established. This project is performed in close collaboration with Dr. Sonja-Verena Albers. One major aim is to further improve and develop the existing expression systems in S. acidocaldarius in order to provide a platform for the expression of (hyper)thermophilic or heat-stable archaeal proteins of biotechnological interest. For expression in S. acidocaldarius the promoter of the maltose binding protein malE is employed. Extensive mutational analysis of the malE promoter in conjunction with insertion of the maltose regulatory protein resulted in significantly increased expression levels compared to the wild type promoter (manuscript in preparation). The improved archaeal expression system is currently used for functional screening of a thermophilic metagenome library for esterase activity. For this reason, an esterase deletion mutant of S. acidocaldarius was constructed. Comparative expression studies will be performed in different expression hosts: S. acidocaldarius, P. putida, R. capsulatus, T. thermopilus and P. antarctica.
.
Person in charge: Julia Kort (M.Sc.)
Email: [email protected] Phone: 0201-183 7065
Electron microcroscopy picture of S. acidocaldarius (S.-V. Albers).
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Study of the microbiological effect of silver nanoparticles and silver-doped CaP nanoparticles. Synthesis, characterization and biological effects
Funded by: Mercator Research Center (MERCUR) Project period: 01.03.2011 until 29.02.2012 Collaborative Project - Project Partners: Prof. M. Epple, Inorganic Chemistry and Center for Nanointegration, University Duisburg-Essen; Prof. M. Köller, Bergmannsheil University Hospital/Surgical Research, Ruhr-University of Bochum; Dr. H. Kuhn, Theoretical Chemistry, University Duisburg-Essen; Prof. H. Rehage, Physical Chemistry II, Technical University of Dortmund Silver-containing chemical materials are increasingly used in various areas (textile industry, medical equipment, cosmetics, etc.) as antimicrobial agent, although the biological effect of silver (colloidal silver, silver ions) is not fully understood. The motivation for this project is to comparatively investigate the antimicrobial effect of ionic silver, silver nanoparticles and silver doped calcium phosphate nanoparticles on eukaryal and bacterial cells.
Contribution of Biofilm-Centre to the consortium: Measurement of bacteriostatic and bactericidal effect of nanosilver and silver-doped nanoparticles. Silver nanparticles and silver doped CaP nanoparticles were synthesized and characterized (Epple). In order to study the silver effect on bacterial cells two model organisms E. coli (Siebers) and S. aureus (Köller) were chosen. To improve comparability to eukaryotic cells, the bacterial cells were grown in cell culture media (RPMI + FCS). The biological effect of the silver materials was investigated by determination of the bacteriostatic and bactericidal concentrations of the respective compounds and the results were compared to those obtained for eukaryotic cell cultures (Köller). Surprisingly the study revealed a rather small therapeutic window for silver in nanoparticulate or ionic form towards mammalian and bacterial cell raising concerns about its widespread application (Peetsch et al. 2012, Greulich et al. 2012).
Person in charge: Dieter Braun
For further information: Email: bettina [email protected] Scanning electron micrographs Phone: 0201-183-7061 of silver ion-doped calcium phosphate nanoparticles. (A. Peetsch)
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Hot biofilms: Biofilm formation and EPS synthesis of the thermoacidophilic Archaeon Sulfolobus acidocaldarius
Funded by: University Duisburg-Essen Project period: October 2009 - Collaborative Project - Project Partners: Dr. Sonja-Verena Albers, Max-Planck-Institute for terrestrial Microbiology, Marburg, Germany
Biofilms represent the common mode of life for more than 99 % of all bacteria in nature.
Bacterial as well as eukaryotic biofilms and their extracellular polymeric substances (EPS)
have been intensively studied because of their ecological importance, application in
biotechnology, but also due to their potential role in human infections and function as
environmental reservoirs for pathogens.
Members of the third domain of life, Archaea, have gained special research interest due to
their adaption to extreme environments. However, the study of biofilm formation and
synthesis of EPS in Archaea has been neglected so far.
The major aim of our study is to unravel biofilm formation as well as EPS synthesis and
composition in the model strain Sulfolobus acidocaldarius. S. acidocaldarius, a
thermoacidophilic, aerobic member of the Crenarchaeota, was first isolated from acid hot
springs at Yellowstone National Park in 1972. Optimal growth is achieved at 78°C at a pH of
2.5-3.5.
In the course of the ongoing project growth conditions for biofilm formation were optimized in
order to characterize S. acidocaldarius biofilms using microscopic methods and their
quantitative and qualitative EPS composition. With the establishment of a suitable EPS
isolation method, EPS from different gene deletion mutants of S. acidocaldarius as well as of
S. solfataricus were successfully isolated and quantified (Koerdt et al., 2012). The method is
currently used to determine the effect of deletion of certain glycosyltransferases on the EPS
composition.
Person in charge: Silke Jachlewski
Email: [email protected] Phone: 0049 (0) 201 183 7067
S. acidocaldarius biofilm on a polycarbonate membrane filter
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Hot signal transduction in Sulfolobus: Regulatory protein phosphorylation in thermoacidophilic Archaea
Funded by: DFG/ University of Duisburg Essen Project period: 01.07.2011 – 01.07.2014 Collaborative Project - Project Partners: Dr. Phillip C. Wright, University of Sheffield, UK; Dr. Sonja-Verena Albers, Max-Plank-Institute for terrestial Microbiology, Marburg, Germany In recent years, it is becoming evident that posttranslational protein modifications like phosporylation, methylation and many more have a major impact on the activity and regulation of proteins and regulatory networks. In Eukaryotes serine/threonine and tyrosine kinases play a major role in such regulatory networks by transducing signals and amplifying them. In the third domain of life, the Archaea, the common bacterial two component systems are largely absent, but all archaea contain a set of eukaryotic type like protein kinases (ePKs) and protein phosphatases. However, only very few of these proteins have been studied and virtually no signal cascade has been unraveled. Model organisms of this study are the thermoacidophilic Crenarchaeota Sulfolobus solfataricus and S. acidocaldarius.
Contribution of Biofilm Centre to the consortium: Investigation of the phospho-proteome, protein kinases, protein phosphatases and predicted target enzymes of the central carbohydrate metabolism and stress reponse
The aim of our study is to elucidate the function of archaeal ePKs and phosphatases in Sulfolobus by the use of classical biochemistry, genetics and proteomics (e.g. enzyme characterization, phospho-proteome analysis, construction of knock-out mutants). The phosphoproteome of Sulfolobus solfataricus was analyzed in respect to the offered carbon source glucose vs tryptone revealing an complex regulation pattern (Esser et al., 2012). The role of RIO kinases in Archaea was investigated using bioinformatics analysis providing some first hints for the possible function and putative target proteins in Archaea (Esser & Siebers 2013). Furthermore, three aldehyde dehydrogenases, which are targeted by reversible protein phosphorylation, from S. solfataricus were characterized in detail (Esser et al., 2013). In vivo and in vitro experiments will complement these approaches with a special focus on central carbon metabolism and stress response in the Siebers group. This work will contribute to unravel signal transduction in the third domain of life and to enhance understanding of the more complex signal transduction processes in Eukaryotes.
Person in charge: Dominik Esser (M. Sc.)
Email: [email protected] Electron microscopic image of Phone : 0049 (0) 201-183-7065 Sulfolobus acidocaldarius cells
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Trehalose as stress response in (hyper-)thermophilic Archaea
Funded by: University of Duisburg-Essen Project period: 01.11.2008 - Collaborative Project - Project Partners: Dr. Sonja Verena Albers, Max-Planck Institute for terrestrial Microbiology, Marburg; Dr. Andrei Lupas, Max-Planck Institute for Developmental Biology, Tübingen
The multifunctional disaccharide trehalose is a widespread non-reducing molecule,
consisting of two -1,1 linked glycosyl-glucose molecules. It occurs in members of the three domains of life and plays a major role as carbon source but it is also shown to function as compatible solute in Eukaryotes and Bacteria. Trehalose protects the cell against a wide range of different stress conditions like heat, cold or high osmolarity or desiccation. Still its function in Archaea is unknown.
From comparative genomic and biochemical analysis five pathways of trehalose synthesis were identified in Archaea: (i) the two step trehalose-6-phosphate synthase (TPS) / trehalose-6-phosphatase (TPP) pathway (OtsA/OtsB), (ii) the two step maltooligosyl-trehalose synthase / trehalohydrolase pathway (TreY/TreZ), (iii) the reversible and the (iv) irreversible trehalose glycosyltransferring synthase pathway (TreT) and (v) the trehalose synthase pathway (TreS).
Our main aim is to unravel the function of trehalose in Archaea. Therefore three Crenarchaeota, which differ in pathway composition, the hyperthermophile Thermoproteus tenax harbouring the unidirectional TreT and the TPSP pathway and the thermoacidophiles Sulfolobus acidocaldarius and S. solfataricus possessing the TreT and the TreY/TreZ pathway, were selected. Different deletion mutants of S. acidocaldarius were established and the phenotype was analysed by growth studies. The results point to a protective role of trehalose during osmotic stress. Additionally protein interaction studies were performed for TreT and OrfY (the encoding genes form an operon) using the yeast two hybrid system.
Finally, the first trehalose synthase complex in Archaea was observed in T. tenax and analyzed via biochemical and molecular biological techniques. This complex consists of the bifunctional TPSP and a glycosyl transferase (GT) which activates the TPSP (manuscript submitted).
Person in charge: Anna Hagemann
Email: [email protected] Phone: 0049 (0) 201-1837065
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Functional analysis of multiple general transcription factors in the crenarchaeal model organism Sulfolobus acidocaldarius
Funded by: Deutsche Forschungsgemeinschaft Project period: 02.11.2009 until 30.09.2015 Collaborative Project - Project Partners: GRK1431 (Transcription, Chromatin Structure and DNA Repair in Development and Differentiation) Homepage: http://www.uni-due.de/zmb/studycourses/gk/gk_home.shtml
Archaea exhibit unique features as well as share characteristics with Bacteria and Eukaryotes. Archaea lack a nucleus, they have a relatively small circular chromosome like Bacteria and genes are organized in operon structures. In contrast to that, information processing e.g. replication, transcription and translation are more eukaryal like.
The archaeal transcription machinery encompasses one multi-subunit RNA-Polymerase (RNAP), resembling the RNAP II of Eukaryotes, homologues of the TATA-binding protein (TBP) and Transcription Factor TFIIB (TFB). The current mechanistic understanding of transcription initiation is that TBP binds to the TATA-Box (~ 25 bp upstream of the transcription start site) whereupon TFB binds to the TBP:DNA complex forming sequence specific contacts with a purine-rich TFB-responsive element (BRE). Subsequently the N-terminus of TFB recruits the RNAP to build the ternary pre-initiation complex. RNAP, TBP and TFB are solely sufficient for transcription of archaeal promoters in vitro. Therefore archaeal transcription is generally regarded as a simpler model of the more complex eukaryal processes.
Interestingly, Archaea possess multiple copies of general transcription factors (GTFs) however, the distribution is species-dependent. Whereas the function of multiple GTFs has been addressed in different Euryarchaeota (e.g. P. furiosus, Halobacterium NRC-1) the role in Crenarchaeota is still unclear. In general a function in stress response similar to bacterial sigma factors has been proposed.
The current study focuses on the thermoacidophile Crenarchaote Sulfolobus acidocaldarius which grows optimally at 80°C and a pH of 2-3. This organism possesses three TFBs and one TBP. TFB1 and TFB2 are full-length proteins with an N-terminal Zn-domain and a C-terminal imperfect direct repeat, while TFB3 lacks the B-finger and DNA-binding domain. The TFIIB homologue most often expressed under normal growing conditions is TFB1, although it has been reported that TFB3 is highly upregulated following UV exposure and acts as a co-activator in the presence of TFB1.The aim of this project is to elucidate the function of multiple transcription factors especially of TFB2 by (i) overexpression of recombinant transcription factors, (ii) purification of RNAP and transcription factors via integrated genomic tags, (iii) knock-out mutant analysis and (iv) protein-interaction studies with RNAP and transcription factors using the yeast two-hybrid approach. Promoter activity of the transcription factors will be tested by reporter gene assays.
Person in charge: Bernadette Rauch
Email: [email protected] Phone: 0049 (0) 201-1837067
Transcription initiation complex of Archaea
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Bachelor- and Master Thesises in 2012
Bachelor Theses accomplished 2012 within or supervised by the Biofilm Centre
Name Title Home Supervisor
Host Supervisor
Host Institution
Anne Ackmann
Methodenvergleich von Verfahren zur Bestimmung der Gesamtzellzahl in Biofilmen
Prof. Dr. Flemming
Dr. Jürgen Gebel
Uni Bonn
Katja Bajohr
Photobiologische Charakterisierung von Mutanten einer Cyclase aus Microcoleus chthonoplstes
Prof. Dr. Siebers
Prof. Dr. Gärtner
MPI Mülheim
Stephan Christel
Bioleaching of Chalcopyrite: Metal Release and Molecular Phylogeny
Prof. Dr. Sand Dr. Mark Dopson
Uni Linnaeus, Sweden
Jennifer Dobbener
Methodenvergleich der ICP-.OES zur ICP-MS an ausgewählten Proben und Ermittlung der relevanten statistischen Kenndaten
Prof. Dr. Schmidt
Dr. Lars Füchtjohann
HuK Umweltlabor
Martin Hachen
Untersuchungen zur Detektion extrazellulärer polymerer Substanzen (EPS) in Biofilmen mittels fluoreszenzmarkierter Lektine
Prof. Dr. Flemming
Dr. Ronald Schade
IBA, Heiligenstadt
Birk Hahne
Determination of the density from different Bacteria regarding phosphate accumulating bacteria
Prof. Dr. Flemming
Prof. Dr. Kartik Chandran
University of Columbia, USA
Patricia Julia Kahl
Einnistung von Legionellen in autochthonen Trinkwasserbiofilmen
Prof. Dr. Flemming
Dr. Jürgen Gebel
Uni Bonn
Senem Kara
Untersuchung von Quecksilberverbindungen mittels Ionenmobilitätsspektrometrie (IMS) mit vorgeschalteter Thermodesoptionseinheit
Prof. Dr. Telgheder
Dr. Holger Krohn
Uni Duisburg-Essen
Tanja Knipping
The role of archaeal EPS in the formation of microbial glass and the preservation of active chemical signals during desiccation
Prof. Dr. Flemming
Prof. Dr. Alan W. Decho
University of South Carolina, USA
Oliver Knoop
Microbial Fuel Cells Prof. Dr. Flemming
Dr. Ioannis A. Ieropoulos
Uni Bristol UK
36
Sarah Küpper
Photometrische Bestimmung von Arsen in Wasser mittels Goldnanopartikeln- Untersuchungen über die Verteilung von Ligand und Arsen in der Goldnanopartikel-Dispersion zur Verbesserung der Bestimmungsgrenze
Prof. Dr. Schmidt
Dr. Achim Rübel
IWW Mülheim
Kristina Lang
Development and validation of a practicable method for the determination of natural and synthetic steroids in water by using the online SPE-HPLC-MS/MS
Prof. Dr. Telgheder
Dr. Friedrich Werres
IWW Mülheim
Markus Libera
Bewertung nicht-thermischer Plasmen zur Entfernung organischer Spurenstoffe Unterwasser
Prof. Dr. Telgheder
Dr. Holger Krohn
Uni Duisburg-Essen
Jonas Mechelke
Charakterisierung von neuen GAF Domänen
Prof. Dr. Siebers
Prof. Dr. Gärtner
MPI Mülheim
Thomas Meyer
Examination of Sulfolobus acidocaldarius for the ability of excpression of Extremozymes
Prof. Dr. Siebers
Dr. Christopher Bräsen
Uni Duisburg-Essen
Min Suk Nauendorf
Entwicklung einer Handlungsempfehlung für den Einsatz des Tropfkörperverfahrens zur Wasseraufbereitung und Wiederverwendung für die nachhaltige Wasserversorgung Afrikas
Prof. Dr. Denecke
Dr. Gerhard Schories
Ttz Bremerhaven
Martin Offermann
Biofilm permeability in RO biofouling and effect of NO treatment
Prof. Dr. Flemming
Prof. Dr. Anthony Fane
Nanyang University Singapore
Mark Pannekens
Biofilmbildung von schwefelreduzierenden Bakterien auf Cyclodextrin beschichteten Stahloberflächen
Prof. Dr. Sand Dr. Mario Vera
Uni Duisburg-Essen
Ines Schäfer Analyse von organischen Spurenstoffen mittels ESI-FAIMS
Prof. Dr. Telgheder
Dr. Holger Krohn
Uni Duisburg-Essen
Gerhard Schertzinger
Multielementanalysen biologischer Proben: Validierung und Fallstudie
Prof. Dr. Schmidt
Prof. Dr. B. Sures
Uni Duisburg-Essen
Sarah Schroth
Untersuchung der Sorption flüchtiger, nicht-ionischer Verbindungen an mehrwandigen Kohlenstoff-Nanoröhren mittels HS-GC-MS Methode
Prof. Dr. Schmidt
Prof. Dr. Telgheder
Uni Duisburg-Essen
37
Frederick Schulz
Transfomationsprozesse anorganischer Schwefelverbindungen in Pflanzenkläranlagen
Prof. Dr. Flemming
Dr. Peter Kuschk
Helmholtz-Zentrum f. Umweltforschung, Leipzig
Philipp Schumann
Presence and persistence of bacterial pathogen Escherichia coli O157: H7 within pilot laboratory scale constructed wetland wastewater treatment systems
Prof. Dr. Flemming
Rachel van Kempen-Fryling
Center for Biofilm Eng USA
Martin Sobotivic
Die Autoxidation und der Einfluss verschiedener Liganden bei der Fenton-Reaktion
Prof. Dr. Schmidt
Prof. Dr. Clemens von Sonntag
Uni Duisburg-Essen
Viola Tacke Neuentwicklung GC-MS-Methode (Headspace) für Siloxane in Gasphase
Prof. Dr. Schmidt
Ralf Hövermann
Wupperverband Wuppertal
Alexander Timm
The analysis of toxic metal ions in samples using modified gold nanoparticles
Prof. Dr. Schmidt
Prof. Dr. Fuangfa Unob
Chulalongkorn University Bangkok
Sebastian Vetter
Fate of extracellular polymeric substances (EPS) after nitric oxide (NO)-induced biofilm dispersal in water filtration systems
Prof. Dr. Flemming
Dr. Nicolas Barraud
University of New South Wales Australia
Mara Werner
Effect of cyclodextrin coating on attachment and corrosiveness of sulfate-reducing or manganese oxidizing microorganisms to stainless steel or carbon steel
Prof. Dr. Sand Prof. Dr. Wolfram Fürbeth
DECHEMA Frankfurt
Fabian Wirth
Evaluierung eines Biosensorsystems zur Bestimmung der Estrogenität von Kläranlagenabläufen vor und nach der Ozonung
Prof. Dr. Flemming
Dr. Jochen Türk
IUTA Duisburg
Sermin Yasar
Bedeutung von Nährstoffsituation, Temperatur und Acanthamoeba castellanii auf die Kultivierbarkeit von Legionella pneumophila
Prof. Dr. Flemming
Dr. Gabriela Schaule
IWW Mülheim
38
Master Theses accomplished 2012 within or supervised by the Biofilm Centre
Name Title Home Supervisor
Host Supervisor
Host Institution
Rani Bakkour
Oxidative degradation of pollutants in sulfate radical-based oxidation in water treatment – mechanistic aspects, efficiency and by-product formation
Prof. Dr. Schmidt
Prof. Dr. Von Sonntag
Uni Duisburg-Essen
Julia Brekenfeld
Degradation of perfluorinated carboxylic acids in the presence of sulfate radicals: degradation products and rate constants
Prof. Dr. Schmidt
Prof. Dr. von Sonntag
Uni Duisburg-Essen
Robert Keith Brown
Microbial fuel cells: Development to prototype status
Prof. Dr. Flemming
Prof. U. Schröder
TU Braunschweig
Jasmine Natalie Hanke
Detection of Escherichia coli in drinking water biofilms by means of culture-independent methods
Prof. Dr. Flemming
Dr. Jost Wingender
Biofilm Centre Germany
Tobias Philipp Häselhoff
Analysis of the behavior of perfluorinated compounds during hydrothermal carbonization
Prof. Dr. Schmidt
Dr. Jochen Türk
IUTA, Duisburg
Ardalan Hashemi ol Hosseini
Characterization of putative temperature sensitive mutants of Legionella pneumophila Corby
Prof. Dr. Siebers
Prof. Dr. Klaus Heuner
RKI Berlin
Heinz-Dietmar Hiegemann
Optimization of deammonificationn by adapted process management
Prof. Dr. Flemming
Prof. Dr. Norbert Jardin
Ruhrverband Essen
Nils Kerlin
Temperatur und pH-Wertkompensation bei der potentiostatischen Onlinemessung von freiem Chlor im Trinkwasser
Prof. Dr. Schmidt
Dr. Wolfgang Babel
Krohne Duisburg
Pascal Kosse
Evaluation of microalgae-bacteria systems for the removal of nutrients in wastewater treatment
Prof. Dr. Flemming
Prof. Dr. Martin Denecke
Uni Duisburg-Essen
Pablo Lores Lareo
Evaluation of different enrichment techniques for the analysis of microbial metabolites by GC-MS/FAIMS in complex matrices
Prof. Dr. Telgheder
Dr. Holger Krohn
Uni Duisburg-Essen
Celine Naderipour
Cross flow experiments for examination of the fouling processes in an anaerobic membrane bioreactor
Prof. Dr. Gimbel
Dipl. Ing. Christoph Brandt (Prof. Dr. Matthias Kraume)
TU Berlin
39
Xochitli Libertad Osorio
Determination of low-molexular mass aldehydes (C1-C10) by headspace in-tube extraction in different packing cardboards used in food products and electro appliances
Prof. Dr. Schmidt
Dr. Maik Jochmann
Uni Duisburg-Essen
Haoyun Pan
Chlorine dioxide sensor evaluation and temperature and pH value compensation in drinking water treatment
Prof. Dr. Schmidt
Dr. Wolfgang Babel
Krohne Duisburg
Marius Paul Rütering
Characterization of Glycolipids by Fermentation of Engineered Strains
Prof. Dr. Siebers
Dr. Martin Schilling
Evonik Essen
Patricia Uloa Almendras
Development and characterization of tissue simulation fluids for MRI
Prof. Dr. Mayer
Prof. Dr. Mark Ladd
Uni Duisburg-Essen
Florian Uteschil
Development of an desolvation chamber for coupling of ESI FAIMS
Prof. Dr. Telgheder
Dr. Holger Krohn
Uni Duisburg-Essen
Sarah Willach
Optimisation and Application of an Analytical Method for the Determination of Adsorbable Organofluorine Compounds (AOF) in the Water Cycle
Prof. Dr. Schmidt
Dr. Frank Thomas Lange
TZW Karlsruhe
40
Publications of Biofilm Centre in 2010
Books
Flemming, H.-C. (Hrsg.): Vermeidung und Sanierung von Trinkwasser-Kontaminationen durch hygienisch relevante Mikroorganismen aus Biofilmen der Hausinstallation „Biofilme in der Hausinstallation“. IWW Schriftenreihe, Mülheim 2010, 258 pp.
Flemming, H.-C., Wingender, J., Szewzyk, U. (eds.): Biofilm Highlights. Springer Int. Heidelberg, New York, 2011, 243 pp
Articles 2010
Balzer, M., Witt, N., Flemming, H.-C., Wingender, J. (2010): Accumulation of fecal indicator bacteria in river biofilms. Wat. Sci. Technol. 61, 1105-1111
Berndt, E., Behnke, S., Dannehl, A., Gajda, A., Wingender, J., Ulbricht, M. (2010) Functional coatings for anti-biofouling applications by surface segregation of block copolymer additives. Polymer 51, 5910-5920
Exner, M., Christiansen, B., Flemming, H.J., Gebel, J., Kistemann, T., Kramer, A., Martiny, M., Mathys, W., Nissing, W., Pleischl, S., Simon, A., Trautmann, M., Zastrow K.D., Engelhart, S. (2010): Pseudomonas aeruginosa – Plädoyer für die Einführung eines technischen Maßnahmewertes in die Novelle der Trinkwasserverordnung. Hyg. Med. 35, 370 – 379
Flemming, H.-C. (2010): Biodeterioration of synthetic materials – a brief review. Mat. Corr. 61, 986-992
Flemming, H.-C., Cloete, T.E. (2010): Environmental impact of controlling biofouling and biocorrosion in cooling water systems. In: Rajagopal, S., Jenner, H.A., Venugopalan, V.P. (eds.): Operational and Environmental Consequences of Large Industrial Cooling Water Systems, 365-380
Flemming, H.-C., Wingender, J. (2010): The Biofilm Matrix. Nat. Rev. Microbiol. 8 623-633
Flemming, H.-C., Wingender, J. (2010): Biofilme – das Gesellschaftsleben der Mikroorganismen. In: Nießner, R. (Hrsg.): Höll, Karl: Wasser - Nutzung im Kreislauf: Hygiene, Analyse und Bewertung, 9. Auflage. De Gruyter, Berlin/New York 2010, pp 660-677
Grobe, S. Schaule, G. Wingender, J., Flemming, H.-C. (2010) Mikrobiologische Kontaminationen im Trinkwasser - Ursachenermittlung. Energie Wasser-Praxis 61 (4), 18-21
Melo, L., Flemming, H.-C. (2010): Mechanistic Aspects of Heat Exchanger and Membrane Biofouling and Prevention. In: Amjad, Z. (ed.): The science and technology of industrial water treatment. Taylor and Francis, London, pp. 365-380
Moritz, M., Flemming, H.-C., Wingender, J. (2010): Integration of Pseudomonas aeruginosa and Legionella pneumophila in drinking water biofilms grown on domestic plumbing materials. Int. J. Hyg. Environ. Health 213, 190-197
Noël, N., Florian, B., Sand, W. (2010): AFM & EFM study on attachment of acidophilic leaching organisms. Hydrometallurgy 104, 370–375
Roeder, R.S., Heeg, K., Tarne, P., Benölken, J.K., Schaule, G., Bendinger, B., Flemming, H.-C., Szewzyk, U. (2010): Influence of materials, water qualities and disinfection methods on the drinking water biofilm community. Wat. Pract. Technol. 5, 1-12
Stadler, R., Fürbeth, M., Grooters, C., Janosch, A., Kuklinski, A., Sand, W. (2010): Studies on the Application of Microbially Produced Polymeric Substances As Protecting Layers Against Microbially Influenced Corrosion of Iron And Steel. Paper no. 10209, Proceedings of CORROSION 2010, San Antonio, Texas
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Stadler, R., Wie, L., Fürbeth, W., Grooters, M., Kuklinski, A. (2010): Influence of bacterial exopolymers on cell adhesion of Desulfovibrio vulgaris on high alloyed steel: Corrosion inhibition by extracellular polymeric substances (EPS). Materials and Corrosion 61(12): 1008-1016
Tielen, P., Rosenau, F., Wilhelm, S., Jaeger, K.F., Flemming, H.-C., Wingender, J. (2010): Extracellular enzymes affect biofilm formation in Pseudomonas aeruginosa. Microbiology 156, 2239-2252
Zaparty M., D. Esser, S. Gertig, P. Haferkamp, T. Kouril, A. Manica, T. K. Pham, J. Reimann, K. Schreiber, P. Sierocinski, D. Teichmann, M. van Wolferen, M. von Jan, P. Wieloch, S.-V. Albers, A. J. Driessen, H.-P. Klenk, C. Schleper, D. Schomburg, J. van der Oost, P. C. Wright and B. Siebers (2010). “Hot standards” for the thermoacidophilic archaeon Sulfolobus solfataricus. Extremophiles, 14, 119-42, DOI: 10.1007/s00792-009-0280-0
Articles 2011
Bellenberg S., Vera M. & Sand W. (2011) Transcriptomic studies of capsular polysaccharide export systems involved in biofilm formation by Acidithiobacillus ferrooxidans In: Biohydrometallurgy: Biotech key to unlock mineral resources value, Vol. 1., Q. Guanzhou, J. Tao, Q. Wenqing, L. Xueduan, Y. Yu, W. Haidong (eds.), Proceedings of the 19th International Biohydrometallurgy Symposium, pp 460-464, Changsha, China.
Bellenberg S., Leon-Morales F. Holuscha D., Krok B., Vera, M & Sand, W. (2011) A lectin-based approach for visualization of capsular EPS after attachment and biofilm formation by acidophilic leaching bacteria. In: Biohydrometallurgy: Biotech key to unlock mineral resources value, Vol 1, Q. Guanzhou, J. Tao, Q. Wenqing, L. Xueduan, Y. Yu, W. Haidong (eds.), Proceedings of the 19th International Biohydrometallurgy Symposium, pp 362-365, Changsha, China
Dwidjosiswojo, Z., Richards, J., Moritz, M.M., Dopp, E., Flemming, H.-C., Wingender, J. (2011): Influence of copper ions on the viability and cytotoxicity of Pseudomonas aeruginosa under conditions relevant to drinking water. Int. J. Hyg. Environ. Health 214, 485-492
Esser D, T. Kouril, M. Zaparty, P. Sierocinski, P. P. Chan, T. Lowe, J. Van der Oost, S.-V. Albers, D. Schomburg, K. S. Makarova and B. Siebers (2011): Functional curation of the Sulfolobus solfataricus P2 and S. acidocaldarius 98-3 complete genome sequences. Extremophiles 15, 711-2
Flemming, H.-C.: Microbial biofouling – unsolved problems, insufficient approaches and possible solutions. In: Flemming, Wingender, J., H.-C., Szewzyk, U. (eds.): Biofilm Highlights. Springer Int. Heidelberg, New York, 2011, 81-109
Flemming, H.-C. (2011): Wasser und Leben. In: Zellner, R. (Hrsg.): Chemie über den Wolken. Verlag Chemie, 132-140
Flemming, H.-C. (2011): The perfect slime. Coll. Surf. B: Biointerfaces, Colloids and Surfaces B: Biointerfaces 86, 251-259
Flemming, H.-C. (2011): The fate of sins in water. Vom Wasser 109, 15-17
Flemming, H.-C., Bendinger. B., Exner, M., Gebel, J., Kistemann, T., Schaule, G., Wingender, J. (2011): Hygiene in der Trinkwasser-Installation: die letzten Meter bis zum Wasserhahn. 44. Essener Tagung für Wasser- u. Abfallwirtschaft; Gewässerschutz, Wasser, Abwasser,
Florian B., Noël N., Thyssen C., Felschau I & Sand W. (2011) Some quantitative data on bacterial attachment to pyrite. Minerals Engineering 24, 1132–1138
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Haferkamp, P., Kutschki, S., Treichel, J., Hemeda, H., Sewczyk, K., Hoffmann, D., Zaparty, M., Siebers, B. (2011): An additional glucose dehydrogenase from Sulfolobus solfataricus: fine-tuning of sugar degradation? Biochem Soc Trans. 2011 Jan 19;39(1):77-81.
Koerdt, A., Jachlewski, S., Gosh, A., Wingender, J., Siebers, B., Albers, S.-V. (2012): Complementation of Sulfolobus solfataricus PBL2025 with an α-mannosidase: effects on surface attachment and biofilm formation. Extremophiles 16, 115-125
Marrero Coto, Ehrenhofer-Murray AE, Pons T, Siebers, B (2011): Functionals analysis of archaeal MBF 1 by complementation studies in yeast. Biol Direct. 6, 18
Siebers B., G. Raddatz, M. Zaparty, B. Tjaden, S.-V. Albers, S. D. Bell, F. Blombach, A. Kletzin, N. Kyrpides, C. Lanz, A. Plagens, M. Rampp, A. Rosinus, M. von Jan, K. S. Makarova, H.-P. Klenk, S. C. Schuster and R. Hensel (2011): The complete genome sequence of Thermoproteus tenax: A physiologically versatile member of the Crenarchaeota. PLoS ONE 6 (10)
Thyssen C., Vera M., Janosch, C. Spröer C. & Sand, W. (2011) A novel acidophilic γ- proteobacterium for bioleaching. In: Biohydrometallurgy: Biotech key to unlock mineral resources value, Vol 2, Q. Guanzhou, J. Tao, Q. Wenqing, L. Xueduan, Y. Yu, W. Haidong (eds.), Proceedings of the 19th International Biohydrometallurgy Symposium, pp 979-891, Changsha, China.
Vera M., Krok B., Sand W. & Poetsch A. (2011) High throughput proteomic comparison of data for planktonic and biofilm-forming cells of Acidthiobacillus ferrooxidans ATCC 23270 In: Biohydrometallurgy: Biotech key to unlock mineral resources value, Vol 1, Q. Guanzhou, J. Tao, Q. Wenqing, L. Xueduan, Y. Yu, W. Haidong (eds.), Proceedings of the 19th International Biohydrometallurgy Symposium, pp 358-361, Changsha, China
Wingender, J.: Hygienically relevant microorganisms in biofilms of man-made water systems. In: Flemming, Wingender, J., H.-C., Szewzyk, U. (eds.): Biofilm Highlights. Springer Int. Heidelberg, New York, 2011, 189-238
Wingender, J., Flemming, H.-C. (2011): Biofilms in drinking water and their role as reservoir for pathogens. Int. J. Hyg. Environ. Health 214, 417-423
Zaparty, M., and Siebers, B. (2011). Physiology, Metabolism and Enzymology of Thermoacidophiles. In Extremophiles Handbook K. Horikoshi, G. Antranikian, A.T. Bull, F.T. Robb, and K.O. Stetter, eds., pp 601-639, Springer, Heidelberg, Tokyo
Articles 2012
Bellenberg S., Leon-Morales C. F., Sand W. & Vera M. (2012). Visualization of Capsular Polysaccharide production induction in Acidithiobacillus ferrooxidans. Hydrometallurgy. 129-130. 82-89.
Esser D., K. T. Pham, J. Reimann, S.-V. Albers, B. Siebers and P. C. Wright (2012): Change of carbon source causes dramatic effects in the phospho-proteome of the Archaeon Sulfolobus solfataricus. J Proteome Res. 11, 4823-22
Donner, C., Panglisch, S., Grobe, S., Wingender, J. (2012) Strategien zur Anpassung von Versorgungsanlagen an neue Herausforderungen. Energie Wasser-Praxis 63(5), 40-46
Flemming, H.-C. (2012): Mythos Wasser. In: Ingensiep, W., Popp, W. (Hrsg.): Hygiene und Kultur. Bidlo-Verlag Essen, 89-109
Flemming, H.-C. u. Wingender, J. (2012): Die Stärke der Gemeinsamkeit. BioSpektrum 18, 714-716
González A., Bellenberg S., Mamani S., Ruiz L., Echeverría A., Soulère L., Doutheau A., Demergasso C., Sand W., Queneau Y., Vera M. & Guiliani N. (2012) AHL signaling
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molecules w ith a large acyl chain enhance biofilm formation on sulfur and metal sulfides by the bioleaching bacterium Acidithiobacillus ferrooxidans. Appl. Microbiol Biotech DOI 10.1007/s00253-012-4229-3
Greulich C., D. Braun, A. Peetsch, J. Diendorf, B. Siebers, M. Epple, and M. Köller (2012): The toxic effect of silver ions and silver nanoparticles towards bacteria and human cells occurs in the same concentration range. RSC Adv., 2012, DOI: 10.1039/C2RA20684F
Grobe, S., Wagner, J., Schaule, G., Flemming, H.-C., Wingender, J. (2012) Mikrobiologische Trinkwasserqualität – Welche Folgen hat der Klimawandel? Wasser 2012. Kurzreferate der Jahrestagung der Wasserchemischen Gesellschaft, Neu-Ulm, pp. 450-454
Koerdt A., S. Jachlewski, A. Ghosh, J. Wingender, B. Siebers and SV Albers (2012): Complementation of Sulfolobus solfataricus PBL2025 with an α-mannosidase: effects on surface attachment and biofilm formation. Extremophiles 16, 115-25
Kouril T., A. Kolodkin, M. Zaparty, R. Steuer, P. Ruoff, H.-V. Westerhoff, J. Snoep, B. Siebers and SulfoSYS consortium (2012): Sulfolobus Systems Biology: Cool hot design for metabolic pathways. In Systems Biology of Microorganisms, Edited by Wren B & Robertson B, Horizon Scientific Press and Caister Academic Press, Norwich, UK.
Peetsch A., C. Greulich, D. Braun, C. Stroetges, H. Rehage, B. Siebers, M. Köller, and M. Epple (2012): Silver-doped calcium phosphate nanoparticles: Synthesis, characterization, and toxic effects towards mammalian and prokaryotic cells. Colloids and Surfaces B: Biointerfaces, 10.1016/j.colsurfb.2012.09.040
Siebers B. (2012): The third domain of life - Interview with B. Siebers, International Innovation Magazine, Issue Environment, pp. 30-32.
Ulas T., S. A. Riemer, M. Zaparty, B. Siebers, D. Schomburg (2012): Genome-scale reconstruction and analysis of the metabolic network in the hyperthermophilic Archaeon Sulfolobus solfataricus. PLoS ONE 7(8)
Vera M., Krok B., Bellenberg S., Sand W & Poetsch A. (2012) Shotgun proteomics study of early biofilm formation process of Acidithiobacillus ferrooxidans ATCC 23270 on pyrite. Proteomics. 2013 Jan 14. doi: 10.1002/pmic.201200386
Articles 2013
Dreszer, C., Vrouwenvelder, H., Kruithof, J., Paulitzsch, A., Flemming, H.-C. (2013): Hydraulic permeability of fouling biofilms on membranes. J. Mem. Sci, 429 436–447
Esser D., T. Kouril, F. Talfournier, J. Polkowska, T. Schrader, C. Bräsen and B. Siebers (2013): Unraveling the function of paralogs of the aldehyde dehydrogenase super family from Sulfolobus solfataricus. Extremophiles, Jan 8, [Epub ahead of print]
Esser D and B. Siebers (2013): A typical protein kinases of the Rio family in Archaea. Biochem Soc Trans. 41, 399-404
Flemming, H.-C., Bendinger, B., Exner, M., Kistemann, T., Schaule, G., Szewzyk, U., Wingender, J. (2013): The last meters before the tap: where drinking water quality is at risk. In: van der Kooij, D., van der Wielen, P. (eds.): Microbial growth in drinking water distribution systems and tap water installations. IWA Publishing, chapter 8.
Flemming, H.-C. u. Wingender, J. (2013): Wann sind Bakterien wirklich tot? KIT Haustechnik, Sonderheft Trinkwasserhygiene, 8-11
Flemming, H.-C., (2013): The biofilm mode of life. In: Krauss, G.-J., Nies, D. (eds.): Ecological Biochemistry- Environmental and Interspecies Interactions today. VCH Weinheim, in press
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Flemming, H.-C, Meier, M. (2013): Biofouling in paper production. A review. Biofouling, accepted
Kouril T., P. Wieloch, J. Reimann, M. Wagner, M. Zaparty, S.-V. Albers, D. Schomburg, P. Ruoff and B. Siebers (2013): Unraveling the function of the two Entner-Doudoroff branches in the thermoacidophilic Crenarchaeon Sulfolobus solfataricus P2. FEBS J. 280(4):1126-38.
Vera M., Krok B., Bellenberg S., Sand W & Poetsch A. (2013) Shotgun proteomics study of early biofilm formation process of Acidithiobacillus ferrooxidans ATCC 23270 on pyrite. Proteomics. 2013 Jan 14. doi: 10.1002/pmic.201200386
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Invited Lectures
Hans-Curt Flemming:
March 25, 2012: „Mikrobiologie in der Wasserleitung “. Jahreskongress Deutsche Gesellschaft für Krankenhaushygiene, Berlin
April 14, 2012: „Nanosilberpartikel (AgNP) - Wirkmechanismen und Untersuchungen ihrer möglichen Interaktionen mit Geweben, Zellen und Molekülen. Definition ihres relevanten Unverträglichkeitspotentials“. NanoCare and NanoCluster Conference, Frankfurt
April 23, 2012: “Conceiled pathogens in biofilms”. Singapore Center for Life Science and Engineering (SCELSE) Conference, Singapore
June 26, 2012: „Biofouling – unsolved questions, insufficient countermeasures and light at the horizon“.International Congress on Marine Corrosion and Fouling (ICMCF), Seattle, USA
July 7, 2012: “Leben und Tod im Biofilm”. Ludwigsburger Sommerakademie für Zahnärzte, Ludwigsburg
September 23, 2012: „Membrane biofouling and permeation properties of biofilms“, International Workshop on Membranes, Oxford
October 3, 2012: “Biofouling: cases, causes, countermeasures”, Conference on Biofouling, Jeddah, Saudi Arabia
November 21, 2012: Significance of the surface in drinking water distribution networks: A SecurEau perspective. Water Contamination and Emergency Conference, Mülheim
December 13, 2012: “Life and death in biofilms” 5th MicroBio Kosmos Conference, Athens
Wolfgang Sand:
Januar - TU Dresden - Biokorrosion und Bioleaching zwei Seiten einer Medaille?
Maerz - Geokompetenzzentrumn Freiberg - Geobiotechnologie in der DECHEMA
April/Mai - Univ. Baia Mare - Erasmuskurs Biocorrosion
Mai - BAM Berlin - Grenzflaechenprozesse bei der Biolaugung
September - Eurocorr Istanbul - Interfacial processes in Bioleaching and Biocorrosion
Oktober - Geohannover, Hannover - Mechanismen des Bioleaching
Oktober - Freiberg Workshop Molekulare Methoden in der Geobiotechnologie - Molecular processes in Bioleaching
November - Changsha, China - Short course Biocorrosion
Dezember - Amira & CSIRO, Melbourne - Bioleaching and Biocorrosion mechanisms
Bettina Siebers:
January 1, 2013: Lightening Talk: Kick-off Meeting ERASysAPP – Systems Biology Application (Brussels, Belgium, Dr. Klaus-Peter Michel, Projektträger Jülich) „Systems Biology Application – SulfoSYS: Life in hot acid”“
January 15,.2013: Gastseminar Physiologie der Mikroorganismen, Ruhr Universität Bochum, (Bochum, Prof. Dr. Nicole Frankenberg-Dinkel): “Life in the hot lane”: Central carbohydrate metabolism and regulation in hyperthermophilic Archaea”
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January 24,.2013: SulfoSYSBiOTEC Kick-off Meeting (Essen): “Applied Sulfolobus Systems Biology: Exploiting the hot archaeal metabolic potential for Biotechnology”
July 3, 2012: Molecular Biology of Archaea III (MPI für Terrestrische Mikrobiologie, Marburg) “Sulfolobus Systems Biology: Challenges of life at high temperature
September 25, 2012: Vortrag Evonik (Dr. Martin Schilling) “Life at high temperature: Challenge and potential“
October 9, 2012: V Internatonal Symposium on Biochemistry and Molecular Biology (within the VIII International Congress on Chemistry and Chemical Engineering) (Havana, Cuba) “Archaeal carbohydrate metabolism at high temperature”
November 28,.2013: Igniococcus Treffen (Frankfurt): “Ignicoccus hospitalis -Nanoarchaeum equitans-“
Jost Wingender
April 26 and June 14, 2012: Die letzten Meter auf dem Weg zum Wasserhahn: BMBF-Verbundprojekt – Legionellen und Pseudomonas aeruginosa im Biofilm. Pall Webinar
Ph. D.´s in 2012:
Dr. Theresa Kouril: “Challenging metabolic networks at high temperature: The central carbohydrate metabolism of Sulfolobus solfataricus” Supervisors: Prof. Dr. Hans V. Westerhoff Prof. Dr. Bettina Siebers November 6, 2012, Amsterdam
Dr. Witold Michalowski: „Composition, dynamics and function of extracellular polymeric substances in drinking-water biofilms” Supervisors: Prof. Dr. Hans-Curt Flemming Prof. Dr. Ulrich Szewzyk, TU Berlin November 6, 2012, Essen
Bianca Florian: „Investigation of initial attachment and biofilm formation of mesophilic leaching bacteria in pure and mixed cultures and their efficiency of pyrite dissolution” Supervisors: Prof. Dr. Wolfgang Sand Prof. Dr. Hans-Curt Flemming May 11, 2012, Essen