DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

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

Abstract

Recombinant expression of toxic proteins remains a challenging problem. Furthermore, 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.

Authors:
ORCiD logo; ; ;
Publication Date:
Research Org.:
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1618889
Alternate Identifier(s):
OSTI ID: 1357376
Report Number(s):
LLNL-JRNL-725506
Journal ID: ISSN 1475-2859; 71; PII: 685
Grant/Contract Number:  
15-LW-013; AC52-07NA27344
Resource Type:
Published Article
Journal Name:
Microbial Cell Factories
Additional Journal Information:
Journal Name: Microbial Cell Factories Journal Volume: 16 Journal Issue: 1; Journal ID: ISSN 1475-2859
Publisher:
Springer Science + Business Media
Country of Publication:
United Kingdom
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; bacterial microcompartment; BMC; lysis protein E; bacteriophage phiX174; toxic protein expression

Citation Formats

Yung, Mimi C., Bourguet, Feliza A., Carpenter, Timothy S., and Coleman, Matthew A. Re-directing bacterial microcompartment systems to enhance recombinant expression of lysis protein E from bacteriophage ϕX174 in Escherichia coli. United Kingdom: N. p., 2017. Web. doi:10.1186/s12934-017-0685-x.
Yung, Mimi C., Bourguet, Feliza A., Carpenter, Timothy S., & Coleman, Matthew A. Re-directing bacterial microcompartment systems to enhance recombinant expression of lysis protein E from bacteriophage ϕX174 in Escherichia coli. United Kingdom. https://doi.org/10.1186/s12934-017-0685-x
Yung, Mimi C., Bourguet, Feliza A., Carpenter, Timothy S., and Coleman, Matthew A. Wed . "Re-directing bacterial microcompartment systems to enhance recombinant expression of lysis protein E from bacteriophage ϕX174 in Escherichia coli". United Kingdom. https://doi.org/10.1186/s12934-017-0685-x.
@article{osti_1618889,
title = {Re-directing bacterial microcompartment systems to enhance recombinant expression of lysis protein E from bacteriophage ϕX174 in Escherichia coli},
author = {Yung, Mimi C. and Bourguet, Feliza A. and Carpenter, Timothy S. and Coleman, Matthew A.},
abstractNote = {Recombinant expression of toxic proteins remains a challenging problem. Furthermore, 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.},
doi = {10.1186/s12934-017-0685-x},
journal = {Microbial Cell Factories},
number = 1,
volume = 16,
place = {United Kingdom},
year = {Wed Apr 26 00:00:00 EDT 2017},
month = {Wed Apr 26 00:00:00 EDT 2017}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1186/s12934-017-0685-x

Citation Metrics:
Cited by: 16 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Modularity of a carbon-fixing protein organelle
journal, December 2011

  • Bonacci, W.; Teng, P. K.; Afonso, B.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 2
  • DOI: 10.1073/pnas.1108557109

Identification of a Minimal Peptide Tag for in Vivo and in Vitro Loading of Encapsulin
journal, June 2016


Interactions between the termini of lumen enzymes and shell proteins mediate enzyme encapsulation into bacterial microcompartments
journal, August 2012

  • Fan, C.; Cheng, S.; Sinha, S.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 37
  • DOI: 10.1073/pnas.1207516109

Toxic protein expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system
journal, March 2006

  • Giacalone, Matthew J.; Gentile, Angela M.; Lovitt, Brian T.
  • BioTechniques, Vol. 40, Issue 3
  • DOI: 10.2144/000112112

A comprehensive set of plasmids for vanillate- and xylose-inducible gene expression in Caulobacter crescentus
journal, November 2007

  • Thanbichler, M.; Iniesta, A. A.; Shapiro, L.
  • Nucleic Acids Research, Vol. 35, Issue 20
  • DOI: 10.1093/nar/gkm818

Synthetic biology and the development of tools for metabolic engineering
journal, May 2012


Structural analysis and classification of native proteins from E. coli commonly co-purified by immobilised metal affinity chromatography
journal, September 2006

  • Bolanos-Garcia, Victor Martin; Davies, Owen Richard
  • Biochimica et Biophysica Acta (BBA) - General Subjects, Vol. 1760, Issue 9
  • DOI: 10.1016/j.bbagen.2006.03.027

Directed Evolution of a Protein Container
journal, February 2011


Engineered Protein Nano-Compartments for Targeted Enzyme Localization
journal, March 2012


Purification and Functional Characterization of ϕX174 Lysis Protein E
journal, June 2009

  • Zheng, Yi; Struck, Douglas K.; Young, Ry
  • Biochemistry, Vol. 48, Issue 22
  • DOI: 10.1021/bi900469g

Genetic Analysis of MraY Inhibition by the ϕX174 Protein E
journal, September 2008


Short N-terminal sequences package proteins into bacterial microcompartments
journal, March 2010

  • Fan, C.; Cheng, S.; Liu, Y.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 16
  • DOI: 10.1073/pnas.0913199107

Engineering formation of multiple recombinant Eut protein nanocompartments in E. coli
journal, April 2016

  • Held, Mark; Kolb, Alexander; Perdue, Sarah
  • Scientific Reports, Vol. 6, Issue 1
  • DOI: 10.1038/srep24359

Solution Structure of a Bacterial Microcompartment Targeting Peptide and Its Application in the Construction of an Ethanol Bioreactor
journal, November 2013

  • Lawrence, Andrew D.; Frank, Stefanie; Newnham, Sarah
  • ACS Synthetic Biology, Vol. 3, Issue 7
  • DOI: 10.1021/sb4001118

Encapsulation of multiple cargo proteins within recombinant Eut nanocompartments
journal, July 2016

  • Quin, Maureen B.; Perdue, Sarah A.; Hsu, Szu-Yi
  • Applied Microbiology and Biotechnology, Vol. 100, Issue 21
  • DOI: 10.1007/s00253-016-7737-8

Expression of Highly Toxic Genes in E. coli: Special Strategies and Genetic Tools
journal, February 2006


Calculation of protein extinction coefficients from amino acid sequence data
journal, November 1989


Effect of bio-engineering on size, shape, composition and rigidity of bacterial microcompartments
journal, November 2016

  • Mayer, Matthias J.; Juodeikis, Rokas; Brown, Ian R.
  • Scientific Reports, Vol. 6, Issue 1
  • DOI: 10.1038/srep36899

A synthetic system for expression of components of a bacterial microcompartment
journal, September 2013


Genetic evidence that the bacteriophage phi X174 lysis protein inhibits cell wall synthesis
journal, April 2000

  • Bernhardt, T. G.; Roof, W. D.; Young, R.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 8
  • DOI: 10.1073/pnas.97.8.4297

Design Parameters to Control Synthetic Gene Expression in Escherichia coli
journal, September 2009


Works referencing / citing this record:

Bio-engineering of bacterial microcompartments: a mini review
journal, June 2019

  • Planamente, Sara; Frank, Stefanie
  • Biochemical Society Transactions, Vol. 47, Issue 3
  • DOI: 10.1042/bst20170564

A Bacterial Microcompartment Is Used for Choline Fermentation by Escherichia coli 536
journal, March 2018

  • Herring, Taylor I.; Harris, Tiffany N.; Chowdhury, Chiranjit
  • Journal of Bacteriology, Vol. 200, Issue 10
  • DOI: 10.1128/jb.00764-17

Engineering the Bacterial Microcompartment Domain for Molecular Scaffolding Applications
journal, July 2017