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Identification and functional characterization of a type I signal peptidase gene of Bacillus megaterium DSM319

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Abstract

The sipM gene of Bacillus megaterium encoding a type I signal peptidase (SPase) was isolated and structurally characterized. RNA analysis revealed a transcript size in accordance with a bicistronic operon comprising sipM and an adjacent open reading frame. Inactivation of sipM by targeted gene disruption could not be achieved, indicating its essential role for cell viability since there might be no other type I SPase of major importance present in B. megaterium. Plasmid-assisted amplification of the gene resulted in an increase in activity of the heterologous glucanase used as an extracellular reporter, suggesting a potential bottleneck for protein secretion within this species.

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Acknowledgements

This work was supported by grants from the Federal Ministry of Education and Research (BMBF, Bonn-Bad Godesberg, Germany), grant no. 0319179E and 0312613. M.A. Rachman thanks the German Academic Exchange Service (DAAD, Bonn-Bad Godesberg, Germany) for financial support. The authors wish to thank K. Müller (Münster, Germany) for cloning the fragment carrying sipM in partially fulfillment of her diploma thesis, and J. Paluszynski (Münster, Germany) for checking the language.

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  1. M. A. Rachman

    Present address: Center of Assessment and Application of Bioindustry, The Agency of the Assessment and Application of Technology, Jl M.H. Thamrin 8, 2nd Bldg., 15th F, 10340, Jakarta , Indonesia

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

    H. Nahrstedt, K.- D. Wittchen, M. A. Rachman & F. Meinhardt

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  1. H. Nahrstedt
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Correspondence to F. Meinhardt.

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Nahrstedt, H., Wittchen, K.D., Rachman, M.A. et al. Identification and functional characterization of a type I signal peptidase gene of Bacillus megaterium DSM319. Appl Microbiol Biotechnol 64, 243–249 (2004). https://doi.org/10.1007/s00253-003-1469-2

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  • DOI: https://doi.org/10.1007/s00253-003-1469-2

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