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13:51
The MultiBac Protein Complex Production Platform at the EMBL
Published on: July 11, 2013
Intramembrane protease RasP boosts protein production in Bacillus
Jolanda Neef1, Cristina Bongiorni2, Vivianne J Goosens1,3
1Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Hanzeplein 1, P.O. Box 30001, 9700 RB, Groningen, The Netherlands.
Microbial Cell Factories
|April 6, 2017
Summary
The intramembrane protease RasP limits high-level production of secreted enzymes in Bacillus subtilis. Overexpressing RasP enhances enzyme production and suppresses growth defects, indicating its role in proteolytic membrane sanitation for efficient industrial enzyme manufacturing.
Area of Science:
- Microbiology
- Biotechnology
- Enzyme Engineering
Background:
- Bacillus subtilis is a key industrial host for producing secreted enzymes.
- Challenges persist in efficiently secreting heterologous enzymes, particularly at high cell densities.
- Previous efforts focused on signal peptides and protease removal, leaving protein export bottlenecks unclear.
Purpose of the Study:
- To investigate the role of the intramembrane protease RasP in high-level enzyme production in B. subtilis.
- To determine if RasP impacts the secretion efficiency of heterologous enzymes.
Main Methods:
- Gene deletion and overexpression of rasP in B. subtilis.
- Analysis of precursor processing and extracellular enzyme levels.
- Assessment of enzyme production and cell growth under different conditions.
Main Results:
- Deletion of rasP reduced extracellular levels of α-amylases and serine proteases.
- Overexpression of rasP increased production of specific α-amylases and serine proteases by 2.5- to 10-fold.
- RasP overexpression suppressed growth defects associated with high-level enzyme production.
Conclusions:
- The intramembrane protease RasP limits the production of certain difficult-to-produce secreted heterologous enzymes in B. subtilis.
- Proteolytic membrane sanitation by RasP is crucial for effective enzyme production in Bacillus.
- RasP is a key factor for optimizing industrial enzyme manufacturing processes.
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