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Microbial Megaplasmids pp 297–309Cite as

Megaplasmid pKB1 of the Rubber-Degrading Bacterium Gordonia westfalica Strain Kb1

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Part of the book series: Microbiology Monographs ((MICROMONO,volume 11))

Abstract

Sequence analysis of the circular 101,016-bp plasmid pKB1 from the rubber-degrading bacterium Gordonia westfalica strain Kb1 revealed 105 open reading frames (ORFs) which could be assigned to three functional groups (a) replication and partitioning, (b) catabolism, and (c) conjugative transfer. Successful con-jugative transfer of pKB1 demonstrated the functionality of its conjugative transfer genes. The origin of replication of pKB1 was identified and used for construction of two Escherichia coliGordonia shuttle vectors suitable for several Gordonia species and related genera. As expression of the pKB1-encoded cadA in E. coli mediated resistance to cadmium, cadA was used as a pKB1-specific selection marker to monitor transfer of pKB1 by electroporation and conjugation to taxonomically related bacteria, mediating cadmium resistance to a maximal concentration of 800 μ M to the recombinant pKB1-harboring strains. Plasmid pKB1-free mutants of G. westfalica strain Kb1 had lost the ability to use natural rubber (NR) as sole carbon source, thereby suggesting that genes essential for NR degradation are encoded by pKB1. Transcription analysis of pKB1-encoded genes with a putative metabolic function revealed that ORF42 (a putative cytochrome c oxidase) and ORF6 (a putative epoxide hydrolase) are induced in cells of G. westfalica strain Kb1 during growth on NR but not or only slightly on sodium acetate. As genetic engineering of pKB1 in its host G. westfalica strain Kb1 failed due to the lack of an effective gene transfer system for this strain, pKB1 was transferred to genetically approachable strains for engineering and analysis of pKB1-encoded features.

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Acknowledgments

The authors are very grateful to the members of the laboratory, who were engaged in research on degradation of polyisoprenoides in the last 10 years. Research on micro-bial rubber degradation was in the past and is currently supported by grant provided by the Deutsche Bundesstiftung Umwelt (AZ. 13,072) and the Deutsche Forschungsgemeinschaft (Ste 386/10–), respectively.

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  1. Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstrasse 3, Münster, D-48149, Germany

    Daniel Bröker & Alexander Steinbüchel

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  1. Daniel Bröker
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  2. Alexander Steinbüchel
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  1. Institut fuer Biologie, Mikrobiologie, Humboldt-Universität zu Berlin, Chausseestr. 117, Berlin, 10115, Germany

    Edward Schwartz

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Bröker, D., Steinbüchel, A. (2009). Megaplasmid pKB1 of the Rubber-Degrading Bacterium Gordonia westfalica Strain Kb1. In: Schwartz, E. (eds) Microbial Megaplasmids. Microbiology Monographs, vol 11. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-85467-8_14

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