A study of long-term effects on plasmid-containing Escherichia coli in carbon-limited chemostat using 2D-fluorescence spectrofluorimetry
(2006) In Biotechnology Progress 22(4). p.1132-1139- Abstract
- Strain stability of plasmid-containing recombinant organisms is clearly important for industrial applications. Stability is normally assessed by methods such as selective colony forming units or by simply measuring the recombinant product. These methods are typically performed off-line, are time-consuming, and do not give detailed information on the changes in the metabolism. In the current work, long-term stability of a plasmid-containing strain of Escherichia coli (W3110.shik1) capable of shikimic acid overproduction was studied by means of a 2D-fluorescence sensor (BioView) able to emit and detect light in ranges of 260-560 nm and 300600 nm, respectively. Long-term carbon-limited chemostat experiments were made under both selective... (More)
- Strain stability of plasmid-containing recombinant organisms is clearly important for industrial applications. Stability is normally assessed by methods such as selective colony forming units or by simply measuring the recombinant product. These methods are typically performed off-line, are time-consuming, and do not give detailed information on the changes in the metabolism. In the current work, long-term stability of a plasmid-containing strain of Escherichia coli (W3110.shik1) capable of shikimic acid overproduction was studied by means of a 2D-fluorescence sensor (BioView) able to emit and detect light in ranges of 260-560 nm and 300600 nm, respectively. Long-term carbon-limited chemostat experiments were made under both selective (tetracycline-containing medium) and nonselective conditions. It is shown that the fluorescence spectra provide information about metabolic changes at an earlier stage, thereby giving a noninvasive method for monitoring of strain stability. Further, the fluorescence measurements showed that (i) the metabolic changes in the strain W3110.shik1 with time were qualitatively different in selective and nonselective environment, (ii) plasmid recombination resulted primarily in increased biomass yield, and (iii) a change in metabolism probably involving FAD/FMN and pyridoxal-5-P occurred in all experiments. It was concluded that the strain was not stable in any growth condition for more than about 25 growth generations and even less if plasmid recombination took place. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/399224
- author
- Johansson, Louise LU and Lidén, Gunnar LU
- organization
- publishing date
- 2006
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Biotechnology Progress
- volume
- 22
- issue
- 4
- pages
- 1132 - 1139
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000239493200028
- scopus:33747153049
- ISSN
- 1520-6033
- DOI
- 10.1021/bp060061m
- language
- English
- LU publication?
- yes
- id
- 252e0609-07f3-407d-8c67-7c62efd15f15 (old id 399224)
- date added to LUP
- 2016-04-01 16:41:21
- date last changed
- 2023-09-04 23:23:35
@article{252e0609-07f3-407d-8c67-7c62efd15f15,
abstract = {{Strain stability of plasmid-containing recombinant organisms is clearly important for industrial applications. Stability is normally assessed by methods such as selective colony forming units or by simply measuring the recombinant product. These methods are typically performed off-line, are time-consuming, and do not give detailed information on the changes in the metabolism. In the current work, long-term stability of a plasmid-containing strain of Escherichia coli (W3110.shik1) capable of shikimic acid overproduction was studied by means of a 2D-fluorescence sensor (BioView) able to emit and detect light in ranges of 260-560 nm and 300600 nm, respectively. Long-term carbon-limited chemostat experiments were made under both selective (tetracycline-containing medium) and nonselective conditions. It is shown that the fluorescence spectra provide information about metabolic changes at an earlier stage, thereby giving a noninvasive method for monitoring of strain stability. Further, the fluorescence measurements showed that (i) the metabolic changes in the strain W3110.shik1 with time were qualitatively different in selective and nonselective environment, (ii) plasmid recombination resulted primarily in increased biomass yield, and (iii) a change in metabolism probably involving FAD/FMN and pyridoxal-5-P occurred in all experiments. It was concluded that the strain was not stable in any growth condition for more than about 25 growth generations and even less if plasmid recombination took place.}},
author = {{Johansson, Louise and Lidén, Gunnar}},
issn = {{1520-6033}},
language = {{eng}},
number = {{4}},
pages = {{1132--1139}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Biotechnology Progress}},
title = {{A study of long-term effects on plasmid-containing Escherichia coli in carbon-limited chemostat using 2D-fluorescence spectrofluorimetry}},
url = {{http://dx.doi.org/10.1021/bp060061m}},
doi = {{10.1021/bp060061m}},
volume = {{22}},
year = {{2006}},
}