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Strain improvement of Escherichia coli K-12 for recombinant production of deuterated proteins

Kelpšas, Vinardas LU and Wachenfeldt, Claes von LU (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.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Scientific Reports
volume
9
issue
1
article number
17694
publisher
Nature Publishing Group
external identifiers
  • scopus:85075619507
  • pmid:31776414
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
2020-01-13 02:36:28
@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},
}