Re-directing bacterial microcompartment systems to enhance recombinant expression of lysis protein E from bacteriophage φX174 in Escherichia coli

Mimi C. Yung, Feliza A. Bourguet, Timothy S. Carpenter, Matthew A Coleman

Research output: Contribution to journalArticlepeer-review

16 Scopus citations


Background: Recombinant expression of toxic proteins remains a challenging problem. One potential method to shield toxicity and thus improve expression of these proteins is to encapsulate them within protein compartments to sequester them away from their targets. Many bacteria naturally produce so-called bacterial microcompartments (BMCs) in which enzymes comprising a biosynthetic pathway are encapsulated in a proteinaeous shell, which is in part thought to shield the cells from the toxicity of reaction intermediates. As a proof-of-concept, we attempted to encapsulate toxic, lysis protein E (E) from bacteriophage φX174 inside recombinant BMCs to enhance its expression and achieve higher yields during downstream purification. Results: E was fused with various N-terminal BMC targeting tags (PduP-, PduD-, and EutC-tags, 18-20 amino acids) and co-expressed with appropriate BMC shell proteins that associate with the tags and are required to form BMCs. Only BMC targeted E fusions, but not non-tagged E, could be successfully cloned, suggesting that the BMC tags reduce the toxicity of E. A PduP-tagged E system appeared to achieve the highest expression of E. Co-expression of Pdu BMC shell proteins with PduP-E increased its expression by 20-50%. Affinity purification of PduP-E via Ni-NTA in the presence of Empigen BB detergent yielded 270 μg of PduP-E per L of induced culture. Removal of the PduP-tag via proteolysis resulted in a final yield of 200 μg of E per L of induced culture, a nearly order of magnitude (~sevenfold) improvement compared to prior reports. Conclusions: These results demonstrate improved expression of φX174 lysis protein E via re-directed BMC systems and ultimately higher E purification yields. Similar strategies can be used to enhance expression of other toxic proteins in recombinant Escherichia coli systems.

Original languageEnglish (US)
Article number71
JournalMicrobial Cell Factories
Issue number1
StatePublished - Apr 26 2017
Externally publishedYes


  • Bacterial microcompartment
  • Bacteriophage phiX174
  • BMC
  • Lysis protein E
  • Toxic protein expression

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


Dive into the research topics of 'Re-directing bacterial microcompartment systems to enhance recombinant expression of lysis protein E from bacteriophage φX174 in Escherichia coli'. Together they form a unique fingerprint.

Cite this