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Enhanced protein and biochemical production using CRISPRi-based growth switches
- Source :
- Li, S, Jendresen, C B, Grünberger, A, Ronda, C, Ingemann Jensen, S, Noack, S & Nielsen, A T 2016, ' Enhanced protein and biochemical production using CRISPRi-based growth switches ', Metabolic Engineering, vol. 38, pp. 274-284 . https://doi.org/10.1016/j.ymben.2016.09.003
- Publication Year :
- 2016
- Publisher :
- Elsevier BV, 2016.
-
Abstract
- Production of proteins and biochemicals in microbial cell factories is often limited by carbon and energy spent on excess biomass formation. To address this issue, we developed several genetic growth switches based on CRISPR interference technology. We demonstrate that growth of Escherichia coli can be controlled by repressing the DNA replication machinery, by targeting dnaA and oriC, or by blocking nucleotide synthesis through pyrF or thyA. This way, total GFP-protein production could be increased by up to 2.2-fold. Single-cell dynamic tracking in microfluidic systems was used to confirm functionality of the growth switches. Decoupling of growth from production of biochemicals was demonstrated for mevalonate, a precursor for isoprenoid compounds. Mass yield of mevalonate was increased by 41%, and production was maintained for more than 45 h after activation of the pyrF-based growth switch. The developed methods represent a promising approach for increasing production yield and titer for proteins and biochemicals.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Transcriptional Activation
Cell
Bioengineering
Biology
medicine.disease_cause
01 natural sciences
Applied Microbiology and Biotechnology
Metabolic engineering
03 medical and health sciences
Bacterial Proteins
010608 biotechnology
medicine
Protein biosynthesis
Protein production
Growth switch
Clustered Regularly Interspaced Short Palindromic Repeats
Escherichia coli
Synechococcus
CRISPR interference
DNA replication
CRISPRi
DnaA
Carbon
Recombinant Proteins
030104 developmental biology
medicine.anatomical_structure
Biochemistry
Metabolic Engineering
Yield (chemistry)
Mevalonate production
Genes, Switch
Metabolic Networks and Pathways
Biotechnology
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Li, S, Jendresen, C B, Grünberger, A, Ronda, C, Ingemann Jensen, S, Noack, S & Nielsen, A T 2016, ' Enhanced protein and biochemical production using CRISPRi-based growth switches ', Metabolic Engineering, vol. 38, pp. 274-284 . https://doi.org/10.1016/j.ymben.2016.09.003
- Accession number :
- edsair.doi.dedup.....25664d82dfbc91a34022a7f9d0211c4b