4.3 Article

Comparison of different Bacillus subtilis eexpression systems

Journal

RESEARCH IN MICROBIOLOGY
Volume 161, Issue 9, Pages 791-797

Publisher

ELSEVIER
DOI: 10.1016/j.resmic.2010.09.004

Keywords

Bacillus subtilis; Expression systems; SpoIISA; SpoIISB

Categories

Funding

  1. Slovak Academy of Sciences [2/0016/10, 2/0063/10]
  2. Slovak Research and Development Agency [APVT-51-0278, LPP-0218-06]
  3. European Science Foundation [ESF-EC-0106]
  4. Wellcome Trust [082829/Z/07/Z]
  5. Wellcome Trust [082829/Z/07/Z] Funding Source: Wellcome Trust

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Bacillus subtilis is considered to have great potential as a host for the production and secretion of recombinant proteins. Many different expression systems have been developed for B. subtilis. Here we compare two widely used expression systems, the IPTG-inducible derivative of spac system (hyper-spank) and the xylose-inducible (xyl) to the SURE (subtilin-regulated gene expression) system. Western blot analysis of the membrane protein SpoIISA together with its protein partner SpoIISB showed that the highest expression level of this complex is obtained using the SURE system. Measurement of beta-galactosidase activities of the promoter-lacZ fusions in individual expression systems confirmed that the P-spaS promoter of the SURE system is the strongest of those compared, although the induction/repression ratio reached only 1.84. Based on these results, we conclude that the SURE system is the most efficient of these three B. subtilis expression systems in terms of the amount of expressed product. Remarkably, the yield of the SpoIISA-SpoIISB complex obtained from B. subtilis was comparable to that normally obtained from the Escherichia coli arabinose-inducible expression system. (C) 2010 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

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