Fermentation strategy to enhance plasmid stability during the cultivation of Escherichia coli for the production of recombinant levansucrase

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Title
Fermentation strategy to enhance plasmid stability during the cultivation of Escherichia coli for the production of recombinant levansucrase
Author(s)
Chul Ho Kim; Jang Young Lee; Min-Gon Kim; Ki Bang Song; Jeong Woo Seo; Bong Hyun Chung; Soon Jae Chang; Sang Ki Rhee
Bibliographic Citation
Journal of Fermentation Bioengineering, vol. 86, no. 4, pp. 391-394
Publication Year
1998
Abstract
To produce levansucrase, a fructosyltransferase enzyme, in a recombinant Escherichia coli harboring the levU gene of Zymomonas mobilis, fermentation strategies were examined in terms of induction methods and plasmid stability. Although the recombinant levansucrase was induced rapidly by IPTG, high instability of the plasmid and formation of inclusion bodies were observed. Segregational instability was aggravated by the excretion of β-lactamase into the culture broth during subculturing, which caused an overgrowth of plasmid-free cells. A combination of methicillin (2, 6-dimethoxyphenyl- penicillin) and ampicillin to provide selective pressure was effective in preventing the growth of plasmid-free cells. The population of plasmid- harboring cells was maintained above 95% of the total cells for more than 100 generations under this condition. In order to replace IPTG, which is toxic and too expensive for use in a large scale, D-lactose was tested and found to be favorable as an alternative inducer.
Keyword
LevansucraseZymomonas mobilisplasmid stabilityinduction methods
ISSN
0385-6380
Publisher
Soc Bioscience Bioengineering Japan
DOI
http://dx.doi.org/10.1016/S0922-338X(99)89010-2
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
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