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Engineering formation of multiple recombinant Eut protein nanocompartments in E. coli.
- Source :
-
Scientific reports [Sci Rep] 2016 Apr 11; Vol. 6, pp. 24359. Date of Electronic Publication: 2016 Apr 11. - Publication Year :
- 2016
-
Abstract
- Compartmentalization of designed metabolic pathways within protein based nanocompartments has the potential to increase reaction efficiency in multi-step biosynthetic reactions. We previously demonstrated proof-of-concept of this aim by targeting a functional enzyme to single cellular protein nanocompartments, which were formed upon recombinant expression of the Salmonella enterica LT2 ethanolamine utilization bacterial microcompartment shell proteins EutS or EutSMNLK in Escherichia coli. To optimize this system, increasing overall encapsulated enzyme reaction efficiency, factor(s) required for the production of more than one nanocompartment per cell must be identified. In this work we report that the cupin domain protein EutQ is required for assembly of more than one nanocompartment per cell. Overexpression of EutQ results in multiple nanocompartment assembly in our recombinant system. EutQ specifically interacts with the shell protein EutM in vitro via electrostatic interactions with the putative cytosolic face of EutM. These findings lead to the theory that EutQ could facilitate multiple nanocompartment biogenesis by serving as an assembly hub for shell proteins. This work offers insights into the biogenesis of Eut bacterial microcompartments, and also provides an improved platform for the production of protein based nanocompartments for targeted encapsulation of enzyme pathways.
- Subjects :
- Bacterial Proteins chemistry
Bacterial Proteins genetics
Escherichia coli ultrastructure
Ethanolamine pharmacology
Genetic Engineering
Metabolic Networks and Pathways drug effects
Microscopy, Electron, Transmission
Microscopy, Fluorescence
Plasmids genetics
Plasmids metabolism
Protein Structure, Tertiary
Recombinant Proteins biosynthesis
Recombinant Proteins chemistry
Recombinant Proteins genetics
Salmonella enterica metabolism
Time-Lapse Imaging
Bacterial Proteins metabolism
Escherichia coli metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 6
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 27063436
- Full Text :
- https://doi.org/10.1038/srep24359