Enhanced protein and biochemical production using CRISPRi-based growth switches

Songyuan Li, Christian Bille Jendresen, Alexander Grünberger, Carlotta Ronda, Sheila Ingemann Jensen, Stephan Noack, Alex Toftgaard Nielsen

Research output: Contribution to journalJournal articleResearchpeer-review

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.
Original languageEnglish
JournalMetabolic Engineering
Volume38
Pages (from-to)274-284
Number of pages11
ISSN1096-7176
DOIs
Publication statusPublished - 2016

Keywords

  • CRISPRi
  • Growth switch
  • Metabolic engineering
  • Mevalonate production
  • Protein production

Fingerprint Dive into the research topics of 'Enhanced protein and biochemical production using CRISPRi-based growth switches'. Together they form a unique fingerprint.

Cite this