Journal
METABOLIC ENGINEERING
Volume 38, Issue -, Pages 274-284Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2016.09.003
Keywords
Growth switch; CRISPRi; Metabolic engineering; Mevalonate production; Protein production
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Funding
- Novo Nordisk Foundation
- People Programme (Marie Curie Actions) of the European Union Seventh Framework Programme FP7-People-ITN [317058]
- NNF Center for Biosustainability [Bacterial Cell Factory Optimization] Funding Source: researchfish
- Novo Nordisk Fonden [NNF15OC0015246, NNF10CC1016517] Funding Source: researchfish
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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.
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