| Autor: |
Kouwen TR; Department of Medical Microbiology, University Medical Center Groningen, 9700 RB Groningen, The Netherlands., Nielsen AK, Denham EL, Dubois JY, Dorenbos R, Rasmussen MD, Quax WJ, Freudl R, van Dijl JM |
| Jazyk: |
angličtina |
| Zdroj: |
Applied and environmental microbiology [Appl Environ Microbiol] 2010 Feb; Vol. 76 (3), pp. 659-69. Date of Electronic Publication: 2009 Nov 30. |
| DOI: |
10.1128/AEM.01671-09 |
| Abstrakt: |
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 (pre(lip)) and pro (pro(lip)) 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 pro(lip)-peptide, in combination with the lipase signal peptide (pre(lip)), contributes significantly to the efficient secretion of PhoA by B. subtilis and that pre(lip) 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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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