Strain improvement of Escherichia coli K-12 for recombinant production of deuterated proteins
(2019) In Scientific Reports 9(1).- Abstract
Deuterium isotope labelling is important for structural biology methods such as neutron protein crystallography, nuclear magnetic resonance and small angle neutron scattering studies of proteins. Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affect growth of cells. The generation time for Escherichia coli K-12 in deuterated medium is substantially increased compared to cells grown in hydrogenated (protiated) medium. By using a mutagenesis plasmid based approach we have isolated an E. coli strain derived from E. coli K-12 substrain MG1655 that show increased fitness in deuterium based growth media, without general adaptation to media components. By whole-genome sequencing we... (More)
Deuterium isotope labelling is important for structural biology methods such as neutron protein crystallography, nuclear magnetic resonance and small angle neutron scattering studies of proteins. Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affect growth of cells. The generation time for Escherichia coli K-12 in deuterated medium is substantially increased compared to cells grown in hydrogenated (protiated) medium. By using a mutagenesis plasmid based approach we have isolated an E. coli strain derived from E. coli K-12 substrain MG1655 that show increased fitness in deuterium based growth media, without general adaptation to media components. By whole-genome sequencing we identified the genomic changes in the obtained strain and show that it can be used for recombinant production of perdeuterated proteins in amounts typically needed for structural biology studies.
(Less)
- author
- Kelpšas, Vinardas LU and Wachenfeldt, Claes von LU
- organization
- publishing date
- 2019
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Scientific Reports
- volume
- 9
- issue
- 1
- article number
- 17694
- publisher
- Nature Publishing Group
- external identifiers
-
- pmid:31776414
- scopus:85075619507
- ISSN
- 2045-2322
- DOI
- 10.1038/s41598-019-54196-w
- language
- English
- LU publication?
- yes
- id
- d0d17218-f5f4-45b1-8149-f7396918a18c
- date added to LUP
- 2019-12-16 10:44:58
- date last changed
- 2024-04-02 23:23:55
@article{d0d17218-f5f4-45b1-8149-f7396918a18c,
abstract = {{<p>Deuterium isotope labelling is important for structural biology methods such as neutron protein crystallography, nuclear magnetic resonance and small angle neutron scattering studies of proteins. Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affect growth of cells. The generation time for Escherichia coli K-12 in deuterated medium is substantially increased compared to cells grown in hydrogenated (protiated) medium. By using a mutagenesis plasmid based approach we have isolated an E. coli strain derived from E. coli K-12 substrain MG1655 that show increased fitness in deuterium based growth media, without general adaptation to media components. By whole-genome sequencing we identified the genomic changes in the obtained strain and show that it can be used for recombinant production of perdeuterated proteins in amounts typically needed for structural biology studies.</p>}},
author = {{Kelpšas, Vinardas and Wachenfeldt, Claes von}},
issn = {{2045-2322}},
language = {{eng}},
number = {{1}},
publisher = {{Nature Publishing Group}},
series = {{Scientific Reports}},
title = {{Strain improvement of Escherichia coli K-12 for recombinant production of deuterated proteins}},
url = {{http://dx.doi.org/10.1038/s41598-019-54196-w}},
doi = {{10.1038/s41598-019-54196-w}},
volume = {{9}},
year = {{2019}},
}