Enhanced protein and biochemical production using CRISPRi-based growth switches
Li S, Jendresen CB, Grünberger A, Ronda C, Jensen SI, Noack S, Nielsen AT (2016)
Metabolic Engineering 38: 274-284.
Zeitschriftenaufsatz
| Veröffentlicht | Englisch
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Autor*in
Li, Songyuan;
Jendresen, Christian Bille;
Grünberger, AlexanderUniBi;
Ronda, Carlotta;
Jensen, Sheila Ingemann;
Noack, Stephan;
Nielsen, Alex Toftgaard
Abstract / Bemerkung
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.
Erscheinungsjahr
2016
Zeitschriftentitel
Metabolic Engineering
Band
38
Seite(n)
274-284
ISSN
1096-7176
Page URI
https://pub.uni-bielefeld.de/record/2912739
Zitieren
Li S, Jendresen CB, Grünberger A, et al. Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering. 2016;38:274-284.
Li, S., Jendresen, C. B., Grünberger, A., Ronda, C., Jensen, S. I., Noack, S., & Nielsen, A. T. (2016). Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering, 38, 274-284. doi:10.1016/j.ymben.2016.09.003
Li, Songyuan, Jendresen, Christian Bille, Grünberger, Alexander, Ronda, Carlotta, Jensen, Sheila Ingemann, Noack, Stephan, and Nielsen, Alex Toftgaard. 2016. “Enhanced protein and biochemical production using CRISPRi-based growth switches”. Metabolic Engineering 38: 274-284.
Li, S., Jendresen, C. B., Grünberger, A., Ronda, C., Jensen, S. I., Noack, S., and Nielsen, A. T. (2016). Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering 38, 274-284.
Li, S., et al., 2016. Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering, 38, p 274-284.
S. Li, et al., “Enhanced protein and biochemical production using CRISPRi-based growth switches”, Metabolic Engineering, vol. 38, 2016, pp. 274-284.
Li, S., Jendresen, C.B., Grünberger, A., Ronda, C., Jensen, S.I., Noack, S., Nielsen, A.T.: Enhanced protein and biochemical production using CRISPRi-based growth switches. Metabolic Engineering. 38, 274-284 (2016).
Li, Songyuan, Jendresen, Christian Bille, Grünberger, Alexander, Ronda, Carlotta, Jensen, Sheila Ingemann, Noack, Stephan, and Nielsen, Alex Toftgaard. “Enhanced protein and biochemical production using CRISPRi-based growth switches”. Metabolic Engineering 38 (2016): 274-284.
Daten bereitgestellt von European Bioinformatics Institute (EBI)
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