Enhancing protein perdeuteration by experimental evolution of Escherichia coli K-12 for rapid growth in deuterium-based media
(2021) In Protein Science 30(12). p.2457-2473- Abstract
Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affects growth of cells. Here, we have studied growth of Escherichia coli MG1655, a wild-type laboratory strain of E. coli K-12, in deuterated glycerol minimal medium. The growth rate and final biomass in deuterated medium is substantially reduced compared to cells grown in ordinary medium. By using a multi-generation adaptive laboratory evolution-based approach, we have isolated strains that show increased fitness in deuterium-based growth media. Whole-genome sequencing identified the genomic changes in the obtained strains and show that there are multiple routes to genetic adaptation to growth in deuterium-based media. By screening a... (More)
Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affects growth of cells. Here, we have studied growth of Escherichia coli MG1655, a wild-type laboratory strain of E. coli K-12, in deuterated glycerol minimal medium. The growth rate and final biomass in deuterated medium is substantially reduced compared to cells grown in ordinary medium. By using a multi-generation adaptive laboratory evolution-based approach, we have isolated strains that show increased fitness in deuterium-based growth media. Whole-genome sequencing identified the genomic changes in the obtained strains and show that there are multiple routes to genetic adaptation to growth in deuterium-based media. By screening a collection of single-gene knockouts of nonessential genes, no specific gene was found to be essential for growth in deuterated minimal medium. Deuteration of proteins is of importance for NMR spectroscopy, neutron protein crystallography, neutron reflectometry, and small angle neutron scattering. The laboratory evolved strains, with substantially improved growth rate, were adapted for recombinant protein production by T7 RNA polymerase overexpression systems and shown to be suitable for efficient production of perdeuterated soluble and membrane proteins for structural biology applications.
(Less)
- author
- Kelpšas, Vinardas LU and von Wachenfeldt, Claes LU
- organization
- publishing date
- 2021-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- adaptive experimental evolution, deuteration, isotope labeling, neutron crystallography
- in
- Protein Science
- volume
- 30
- issue
- 12
- pages
- 17 pages
- publisher
- The Protein Society
- external identifiers
-
- scopus:85119456428
- pmid:34655136
- ISSN
- 0961-8368
- DOI
- 10.1002/pro.4206
- language
- English
- LU publication?
- yes
- additional info
- Publisher Copyright: © 2021 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.
- id
- af6a0609-9de0-4db9-bcfc-559df56f09b7
- date added to LUP
- 2021-12-08 21:48:20
- date last changed
- 2024-03-23 15:31:35
@article{af6a0609-9de0-4db9-bcfc-559df56f09b7,
abstract = {{<p>Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affects growth of cells. Here, we have studied growth of Escherichia coli MG1655, a wild-type laboratory strain of E. coli K-12, in deuterated glycerol minimal medium. The growth rate and final biomass in deuterated medium is substantially reduced compared to cells grown in ordinary medium. By using a multi-generation adaptive laboratory evolution-based approach, we have isolated strains that show increased fitness in deuterium-based growth media. Whole-genome sequencing identified the genomic changes in the obtained strains and show that there are multiple routes to genetic adaptation to growth in deuterium-based media. By screening a collection of single-gene knockouts of nonessential genes, no specific gene was found to be essential for growth in deuterated minimal medium. Deuteration of proteins is of importance for NMR spectroscopy, neutron protein crystallography, neutron reflectometry, and small angle neutron scattering. The laboratory evolved strains, with substantially improved growth rate, were adapted for recombinant protein production by T7 RNA polymerase overexpression systems and shown to be suitable for efficient production of perdeuterated soluble and membrane proteins for structural biology applications.</p>}},
author = {{Kelpšas, Vinardas and von Wachenfeldt, Claes}},
issn = {{0961-8368}},
keywords = {{adaptive experimental evolution; deuteration; isotope labeling; neutron crystallography}},
language = {{eng}},
number = {{12}},
pages = {{2457--2473}},
publisher = {{The Protein Society}},
series = {{Protein Science}},
title = {{Enhancing protein perdeuteration by experimental evolution of Escherichia coli K-12 for rapid growth in deuterium-based media}},
url = {{http://dx.doi.org/10.1002/pro.4206}},
doi = {{10.1002/pro.4206}},
volume = {{30}},
year = {{2021}},
}