Genetic or chemical protease inhibition causes significant changes in the Bacillus subtilis exoproteome

Bacillus subtilis is a prolific producer of enzymes and biopharmaceuticals. However, the susceptibility of heterologous proteins to degradation by (extracellular) proteases is a major limitation for use of B. subtilis as a protein cell factory. An increase in protein production levels has previously...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2008-07, Vol.8 (13), p.2704-2713
Main Authors: Westers, Lidia, Westers, Helga, Zanen, Geeske, Antelmann, Haike, Hecker, Michael, Noone, David, Devine, Kevin M, van Dijl, Jan Maarten, Quax, Wim J
Format: Article
Language:English
Subjects:
Alpha-amylase
Analytical, structural and metabolic biochemistry
Bacillus subtilis - enzymology
Bacillus subtilis - genetics
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Biological and medical sciences
Endopeptidases - metabolism
Exoproteome
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Bacterial
Interleukin-3
Miscellaneous
Periplasmic Proteins - genetics
Periplasmic Proteins - metabolism
Protease inhibition
Protease Inhibitors - metabolism
Proteins
Proteome - analysis
Proteomics - methods
Secretome
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Summary:Bacillus subtilis is a prolific producer of enzymes and biopharmaceuticals. However, the susceptibility of heterologous proteins to degradation by (extracellular) proteases is a major limitation for use of B. subtilis as a protein cell factory. An increase in protein production levels has previously been achieved by using either protease-deficient strains or addition of protease inhibitors to B. subtilis cultures. Notably, the effects of genetic and chemical inhibition of proteases have thus far not been compared in a systematic way. In the present studies, we therefore compared the exoproteomes of cells in which extracellular proteases were genetically or chemically inactivated. The results show substantial differences in the relative abundance of various extracellular proteins. Furthermore, a comparison of the effects of genetic and/or chemical protease inhibition on the stress response triggered by (over) production of secreted proteins showed that chemical protease inhibition provoked a genuine secretion stress response. From a physiological point of view, this suggests that the deletion of protease genes is a better way to prevent product degradation than the use of protease inhibitors. Importantly however, studies with human interleukin-3 show that chemical protease inhibition can result in improved production of protease-sensitive secreted proteins even in mutant strains lacking eight extracellular proteases.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200800009