Dynamic feedback regulation for efficient membrane protein production using a small RNA-based genetic circuit in Escherichia coli
- Author
- Chiara Guidi (UGent) , Lien De Wannemaeker (UGent) , Jasmine De Baets (UGent) , Wouter Demeester (UGent) , Jo Maertens (UGent) , Brecht De Paepe (UGent) and Marjan De Mey (UGent)
- Organization
- Project
-
- Syn/SysBio4COS: Combining synthetic and systems biology to unlock the potential of the hexosamine biosynthesis pathway for chitooligosaccharide production
- Development of microbial cell factories for the production of chitoheptaose and -octaose using metabolic and protein engineering: Promising molecules with an underexplored potential for plant, animal and human health
- Disclosing the bacterial domain for industrial biotechnology applications: Cross-Bacterial Expression System (CBES) transplantation to turn Cupriavidus necator into a kaempferolproducing microbial cell factory
- Expanding the genetic circuit repertoire for the optimization of microbial cell factories producing partially acetylated chitooligosaccharides.
- Tuning auto-inducible pathway-independent dynamic switches for the optimisation of the microbial production of partially acetylated chitooligosaccharides
- Synthetic biology and artificial intelligence, mutual learning to advance together and jointly drive industrial biotechnology - accelerating knowledge discovery and strain engineering
- Abstract
- Background: Membrane proteins (MPs) are an important class of molecules with a wide array of cellular functions and are part of many metabolic pathways. Despite their great potential-as therapeutic drug targets or in microbial cell factory optimization-many challenges remain for efficient and functional expression in a host such as Escherichia coli. Results: A dynamically regulated small RNA-based circuit was developed to counter membrane stress caused by overexpression of different MPs. The best performing small RNAs were able to enhance the maximum specific growth rate with 123%. On culture level, the total MP production was increased two-to three-fold compared to a system without dynamic control. This strategy not only improved cell growth and production of the studied MPs, it also suggested the potential use for countering metabolic burden in general. Conclusions: A dynamically regulated feedback circuit was developed that can sense metabolic stress caused by, in casu, the overexpression of an MP and responds to it by balancing the metabolic state of the cell and more specifically by downregulating the expression of the MP of interest. This negative feedback mechanism was established by implementing and optimizing simple-to-use genetic control elements based on post-transcriptional regulation: small non-coding RNAs. In addition to membrane-related stress when the MP accumulated in the cytoplasm as aggregates, the sRNA-based feedback control system was still effective for improving cell growth but resulted in a decreased total protein production. This result suggests promiscuity of the MP sensor for more than solely membrane stress.
- Keywords
- Membrane proteins, Escherichia coli, sRNA-based genetic circuitry, Industrial biotechnology
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-01GPB2WF0KE58HP2ZKH21P5YG0
- MLA
- Guidi, Chiara, et al. “Dynamic Feedback Regulation for Efficient Membrane Protein Production Using a Small RNA-Based Genetic Circuit in Escherichia Coli.” MICROBIAL CELL FACTORIES, vol. 21, no. 1, 2022, doi:10.1186/s12934-022-01983-2.
- APA
- Guidi, C., De Wannemaeker, L., De Baets, J., Demeester, W., Maertens, J., De Paepe, B., & De Mey, M. (2022). Dynamic feedback regulation for efficient membrane protein production using a small RNA-based genetic circuit in Escherichia coli. MICROBIAL CELL FACTORIES, 21(1). https://doi.org/10.1186/s12934-022-01983-2
- Chicago author-date
- Guidi, Chiara, Lien De Wannemaeker, Jasmine De Baets, Wouter Demeester, Jo Maertens, Brecht De Paepe, and Marjan De Mey. 2022. “Dynamic Feedback Regulation for Efficient Membrane Protein Production Using a Small RNA-Based Genetic Circuit in Escherichia Coli.” MICROBIAL CELL FACTORIES 21 (1). https://doi.org/10.1186/s12934-022-01983-2.
- Chicago author-date (all authors)
- Guidi, Chiara, Lien De Wannemaeker, Jasmine De Baets, Wouter Demeester, Jo Maertens, Brecht De Paepe, and Marjan De Mey. 2022. “Dynamic Feedback Regulation for Efficient Membrane Protein Production Using a Small RNA-Based Genetic Circuit in Escherichia Coli.” MICROBIAL CELL FACTORIES 21 (1). doi:10.1186/s12934-022-01983-2.
- Vancouver
- 1.Guidi C, De Wannemaeker L, De Baets J, Demeester W, Maertens J, De Paepe B, et al. Dynamic feedback regulation for efficient membrane protein production using a small RNA-based genetic circuit in Escherichia coli. MICROBIAL CELL FACTORIES. 2022;21(1).
- IEEE
- [1]C. Guidi et al., “Dynamic feedback regulation for efficient membrane protein production using a small RNA-based genetic circuit in Escherichia coli,” MICROBIAL CELL FACTORIES, vol. 21, no. 1, 2022.
@article{01GPB2WF0KE58HP2ZKH21P5YG0,
abstract = {{Background: Membrane proteins (MPs) are an important class of molecules with a wide array of cellular functions and are part of many metabolic pathways. Despite their great potential-as therapeutic drug targets or in microbial cell factory optimization-many challenges remain for efficient and functional expression in a host such as Escherichia coli. Results: A dynamically regulated small RNA-based circuit was developed to counter membrane stress caused by overexpression of different MPs. The best performing small RNAs were able to enhance the maximum specific growth rate with 123%. On culture level, the total MP production was increased two-to three-fold compared to a system without dynamic control. This strategy not only improved cell growth and production of the studied MPs, it also suggested the potential use for countering metabolic burden in general. Conclusions: A dynamically regulated feedback circuit was developed that can sense metabolic stress caused by, in casu, the overexpression of an MP and responds to it by balancing the metabolic state of the cell and more specifically by downregulating the expression of the MP of interest. This negative feedback mechanism was established by implementing and optimizing simple-to-use genetic control elements based on post-transcriptional regulation: small non-coding RNAs. In addition to membrane-related stress when the MP accumulated in the cytoplasm as aggregates, the sRNA-based feedback control system was still effective for improving cell growth but resulted in a decreased total protein production. This result suggests promiscuity of the MP sensor for more than solely membrane stress.}},
articleno = {{260}},
author = {{Guidi, Chiara and De Wannemaeker, Lien and De Baets, Jasmine and Demeester, Wouter and Maertens, Jo and De Paepe, Brecht and De Mey, Marjan}},
issn = {{1475-2859}},
journal = {{MICROBIAL CELL FACTORIES}},
keywords = {{Membrane proteins,Escherichia coli,sRNA-based genetic circuitry,Industrial biotechnology}},
language = {{eng}},
number = {{1}},
pages = {{17}},
title = {{Dynamic feedback regulation for efficient membrane protein production using a small RNA-based genetic circuit in Escherichia coli}},
url = {{http://doi.org/10.1186/s12934-022-01983-2}},
volume = {{21}},
year = {{2022}},
}
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