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Coexpression of the type I signal peptidase gene sipM increases recombinant protein production and export in Bacillus megaterium MS941
Author(s) -
Malten Marco,
Nahrstedt Hannes,
Meinhardt Friedhelm,
Jahn Dieter
Publication year - 2005
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20523
Subject(s) - biology , signal peptide , bacillus megaterium , microbiology and biotechnology , gene , recombinant dna , biochemistry , genetics , bacteria
The removal of the signal peptide from a precursor protein is a crucial step of protein secretion. In order to improve Bacillus megaterium as protein production and secretion host, the influence of homologous type I signal peptidase SipM overproduction on recombinant Leuconostoc mesenteroides dextransucrase DsrS synthesis and export was investigated. The dsrS gene was integrated as a single copy into the chromosomal bgaM locus encoding β‐galactosidase. Desired clones were identified by blue‐white selection. In this strain, the expression of sipM from a multicopy plasmid using its own promoter increased the amount of secreted DsrS 3.7‐fold. This increase in protein secretion by SipM overproduction was next transferred to a high level DsrS production strain using a multicopy plasmid encoding sipM with its natural promoter and dsrS under control of a strong xylose‐inducible promoter. No further increase in DsrS export were observed when this vector was carrying two sipM copies. Similarly, bicistronic sipM and dsrS high level expression did not enhance DsrS secretion, indicating the natural limitation of the approach. Interestingly, SipM‐enhanced DsrS secretion also resulted in an overall increase of DsrS production. © 2005 Wiley Periodicals, Inc.