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Contributions of the pre- and pro-regions of a staphylococcus hyicus lipase to secretion of a heterologous protein by bacillus subtilis
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Kouwen, T. R. H. M., Nielsen, A. K., Denham, Emma, Dubois, J. -Y. F., Dorenbos, R., Rasmussen, M. D., Quax, W. J., Freudl, R. and van Dijl, J. M. (2020) Contributions of the pre- and pro-regions of a staphylococcus hyicus lipase to secretion of a heterologous protein by bacillus subtilis. Applied and Environmental Microbiology, 76 (3). pp. 659-669. doi:10.1128/AEM.01671-09 ISSN 0099-2240.
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Official URL: http://dx.doi.org/10.1128/AEM.01671-09
Abstract
Bacillus subtilis is a well-established cell factory for efficient secretion of many biotechnologically relevant enzymes that are naturally produced by it or related organisms. However, the use of B. subtilis as a host for production of heterologous secretory proteins can be complicated by problems related to inefficient translocation of the foreign proteins across the plasma membrane or to inefficient release of the exported proteins from the cell surface into the surrounding medium. Therefore, there is a clear need for tools that allow more efficient membrane targeting, translocation, and release during the production of these proteins. In the present study, we investigated the contributions of the pre (prelip) and pro (prolip) sequences of a Staphylococcus hyicus lipase to secretion of a heterologous protein, the alkaline phosphatase PhoA of Escherichia coli, by B. subtilis. The results indicate that the presence of the prolip-peptide, in combination with the lipase signal peptide (prelip), contributes significantly to the efficient secretion of PhoA by B. subtilis and that prelip directs PhoA secretion more efficiently than the authentic signal peptide of PhoA. Genome-wide transcriptional analyses of the host cell responses indicate that, under the conditions tested, no known secretion or membrane-cell wall stress responses were provoked by the production of PhoA with any of the pre- and pro-region sequences used. Our data underscore the view that the pre-pro signals of the S. hyicus lipase are very useful tools for secretion of heterologous proteins in B. subtilis.
| Item Type: | Journal Article | ||||||
|---|---|---|---|---|---|---|---|
| Divisions: | Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Microbiology & Infection Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School |
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| Journal or Publication Title: | Applied and Environmental Microbiology | ||||||
| Publisher: | American Society for Microbiology | ||||||
| ISSN: | 0099-2240 | ||||||
| Official Date: | 17 December 2020 | ||||||
| Dates: |
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| Volume: | 76 | ||||||
| Number: | 3 | ||||||
| Page Range: | pp. 659-669 | ||||||
| DOI: | 10.1128/AEM.01671-09 | ||||||
| Status: | Peer Reviewed | ||||||
| Publication Status: | Published | ||||||
| Access rights to Published version: | Restricted or Subscription Access |
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