Biomineralisation of calcite by bacterial isolates

Keiner, Robert; Frosch, Torsten; Hanf, Stefan; Rusznyak, Anna; Akob, Denise M; Küsel, Kirsten; Popp, Jürgen

Raman Spectroscopy—An Innovative and Versatile Tool To Follow the Respirational Activity and Carbonate Biomineralization of Important Cave Bacteria Journal Article

Analytical Chemistry, 85 (18), pp. 8708–8714, 2013.

Abstract | Links | BibTeX

Rusznyák, Anna; Akob, Denise M; Nietzsche, Sándor; Eusterhues, Karin; Totsche, Kai Uwe; Neu, Thomas R; Frosch, Torsten; Popp, Jürgen; Keiner, Robert; Geletneky, Jörn; Katzschmann, Lutz; Schulze, Ernst-Detlef; Küsel, Kirsten

Calcite Biomineralization by Bacterial Isolates from the Recently Discovered Pristine Karstic Herrenberg Cave Journal Article

Applied and Environmental Microbiology, 78 (4), pp. 1157–1167, 2012.

Abstract | Links | BibTeX

Using stalactite material and fluvial sediments from the Blessberg cave, the diversity and activity of the occuring bacteria and their formation of carbonate minerals were investigated. For this purpose, various techniques such as phylogenetic analyses, bacterial cultivation, electron microscopy, X-ray spectroscopy, confocal laser scanning microscopy, and Raman spectroscopy were used.

Bacteria were found on the surface and inside the stalactites using confocal laser scanning microscopy. Phylogenetic analyses showed that Proteobacteria were most abundant in the bacterial communities on and inside the stalactites and also in the fluvial sediments, but also other groups such as Actinobacteria or FirmicutesFirmicutes Eine artenreiche Gruppe innerhalb der Bakterien. Unterscheiden sich von den Actinobakterien unter anderem durch ihren niedrigeren Gehalt an Nukleinbasen in der Bakterien-DNS.. Many of the detected bacteria have not yet been cultivated.

A total of nine new bacterial cultures were isolated from the cave sediments, growing on alkaline carbonate-precipitating medium and belonging to the bacterial genera Arthrobacter, Flavobacterium, Pseudomonas, Rhodococcus, Serratia and Stenotrophomonas. Of these, the two with the most intense precipitate formation were selected for further research: Arthrobacter sulfonivorans SCM3 and Rhodococcus globerulus SCM4. Both produced extracellular polymeric substances (EPS) and grew as cell aggregates.

Scanning electron microscopy of cell aggregates and extracellular polymeric substance (EPS) of the isolates Arthrobacter sulfonivorans SCM3 and Rhodococcus globerulus SCM4
Scanning electron microscopy of cell aggregates and extracellular polymeric substance (EPS) of the isolates Arthrobacter sulfonivorans SCM3 (left) and Rhodococcus globerulus SCM4 (right) (Image: Elektronenmikroskopisches Zentrum Jena, Sándor Nietzsche and Anna Rusznyak).

The carbonate minerals formed were mixtures of calcite, vaterite, and monohydrocalcite. Arthrobacter sulfonivorans SCM3 formed xenomorphic spherical crystals, and Rhodococcus globerulus SCM4 idiomorphic crystals with rhombohedral morphology.

The biomineralisation process of Arthrobacter sulfonivorans SCM3 was further investigated using a combination of Raman macro- and microspectroscopy to obtain a spatially resolved chemical representation of the different types of calcium carbonate minerals. The cell surface of Arthrobacter sulfonivorans SCM3 served as a nucleus for the biomineralisation of vaterite precipitates. These were initially spherical and then continued to grow as chemically stable, rock-forming calcite crystals with rough edges.

Scanning electron microscopy of precipitates of the isolates Arthrobacter sulfonivorans SCM3 and Rhodococcus globerulus SCM4
Scanning electron microscopy of precipitates of the isolates Arthrobacter sulfonivorans SCM3 (left) and Rhodococcus globerulus SCM4 (right) (Image: Elektronenmikroskopisches Zentrum Jena, Sándor Nietzsche and Anna Rusznyak).
Isolat und Präzipitate (Vorschau)

Diversity and role of cave bacteria in the formation of carbonate minerals

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within the framework of the research project “AquaDiv@Jena”, funded by the ProExcellence Initiative of the Free State of Thuringia, and the Collaborative Research Centre 1076 “AquaDiva ↗“, funded by the DFG

Project Lead

Content

What biodiversity do we find in subsurface habitats such as caves? What role do bacteria play in the Earth’s biogeochemical cycles? Which bacteria are involved in the formation of minerals?

Karst areas are particularly interesting subsurface habitats because they represent one of the most important natural subsurface reservoirs of carbon on Earth. Caves are like a window into these subsurface habitats and allow the exploration of subsurface microbial life. However, it can be difficult to distinguish between microbial communities that are pristine or indigenous and those that have been introduced into caves by animals and humans. The Blessberg cave provides a unique pristine site for the study of active bacterial communities in a karst system.

Using stalactite material and fluvialfluvial Durch fließendes Wasser verursacht; zum Beispiel: "Fluviale Sedimente" = durch fließendes Wasser abgelagertes Material. sediments from the Blessberg cave, the diversity and activity of the occurring bacteria and their formation of carbonate minerals are being investigated. For this purpose, a wide variety of techniques such as phylogenetic analyses, bacterial cultivation, electron microscopy, X-ray spectroscopy, confocal laser scanning microscopy and Raman spectroscopy are applied.

Carbonate precipitates of the bacterial isolate Rhodococcus globerulus SCM4
Carbonate precipitates of the bacterial isolate Rhodococcus globerulus SCM4 from the Blessberg cave. EDX mapping underlain by a gray level scanning electron microscopy image; the false colors red, green, and blue show the local occurrence of the elements carbon, oxygen, and calcium, respectively (Image: Elektronenmikroskopisches Zentrum Jena, Sándor Nietzsche and Anna Rusznyak).

Results

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Friedrich Schiller University Jena, Institute of Biodiversity – Group of Aquatic Geomicrobiology

The Aquatic Geomicrobiology group exists since 2004 and is headed by Prof. Dr. Kirsten Küsel. The group is has a focus on the role of microorganisms that are driving the Earth’s biogeochemical cycles. Here, microbes are also involved in mediating mineral precipitation and dissolution as well as the sorption of organic material. The research covers the areas of biodiversity, microbial interactions, climate change mitigation, mining, and the deep biosphere.

In the context of the research project “AquaDiv@Jena” and the CRC 1076 “AquaDiva” ↗, the diversity and activity of the bacteria contained in the Blessberg cave and their formation of carbonate minerals were investigated.

Contact

Prof. Dr. Kirsten Küsel ↗

Web Site

https://www.geomicrobiology.uni-jena.de/ ↗

Publications

Keiner, Robert; Frosch, Torsten; Hanf, Stefan; Rusznyak, Anna; Akob, Denise M; Küsel, Kirsten; Popp, Jürgen

Raman Spectroscopy—An Innovative and Versatile Tool To Follow the Respirational Activity and Carbonate Biomineralization of Important Cave Bacteria Journal Article

Analytical Chemistry, 85 (18), pp. 8708–8714, 2013.

Abstract | Links | BibTeX

Rusznyák, Anna; Akob, Denise M; Nietzsche, Sándor; Eusterhues, Karin; Totsche, Kai Uwe; Neu, Thomas R; Frosch, Torsten; Popp, Jürgen; Keiner, Robert; Geletneky, Jörn; Katzschmann, Lutz; Schulze, Ernst-Detlef; Küsel, Kirsten

Calcite Biomineralization by Bacterial Isolates from the Recently Discovered Pristine Karstic Herrenberg Cave Journal Article

Applied and Environmental Microbiology, 78 (4), pp. 1157–1167, 2012.

Abstract | Links | BibTeX