Articles & Issues

Language: English

Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received December 25, 2007
Accepted February 27, 2008

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

All issues

A fermentation strategy for production of recombinant protein subjected to plasmid instability

Shu-Jen Chen^† Bo-Shun Ke¹ I-Chung Chiu¹

Department of Chemical and Materials Engineering, National Kaohsiung University of Applied Sciences, No. 415, Chien Kung Road, Kaohsiung 80778, Taiwan ¹Department of Chemical Engineering, National Cheng Kung University, No.1, University Road, Tainan 70101, Taiwan

biochen@cc.kuas.edu.tw

Korean Journal of Chemical Engineering, September 2008, 25(5), 1110-1114(5), 10.1007/s11814-008-0181-4

Download PDF

Abstract

The appearance of plasmid-losing cells in a recombinant Escherichia coli culture was observed when the cell mass became doubled after induction, which corresponded to the timing of cell fission. Accordingly, a two-stage fermentation strategy capable of maintaining plasmid stability without selective pressure in a recombinant E. coli culture was proposed. In the first stage (cell growth stage), a high cell density culture was obtained by incubating the cells in the R medium. In the second stage (producing stage), the cells were devoted to producing the recombinant protein by introducing the fresh LB medium supplemented with isopropyl-β-D-thiogalactopyranoside (IPTG). It was necessary to prevent the doubling in the cell mass after induction; otherwise cell fission would occur and generate plasmid-losing cells. The present strategy is expected to be extensively applicable in recombinant E. coli cultures.

Keywords

Escherichia coli Induction Plasmid Stability Recombinant Two-Stage Culture

References

Seo JH, Bailey JE, Biotechnol. Bioeng., 27, 1668 (1985)
LEE K, CHOI CY, Korean J. Chem. Eng., 4(2), 182 (1987)
Bentley WE, Mirjalili N, Andersen DC, Davis RH, Kompala DS, Biotechnol. Bioeng., 35, 668 (1990)
Shin PK, Koo JH, Lee WJ, Seo JH, Korean J. Chem. Eng., 13(1), 82 (1996)
Kim CH, Lee JY, Kim MG, Song KB, Seo JW, Chung BH, Chang SJ, Rhee SK, J. Ferment. Bioeng., 86(4), 391 (1998)
LEE JH, YOO YJ, PAR KH, Korean J. Chem. Eng., 8(1), 39 (1991)
DiBiasio D, Sardonini C, Ann. NY Acad. Sci., 469, 111 (1986)
De Taxis Du Poet P, Acand Y, Bernier Jr R, Barbotin JN, Thomas D, Appl. Environ. Microbiol., 53, 1548 (1987)
Stephens ML, Christensen C, Lyberatos G, Biotechnol. Prog., 8, 1 (1992)
Weber AE, San KY, Biotechnol. Lett., 10, 531 (1988)
Henry KL, Davis RH, Taylor AL, Biotechnol. Prog., 6, 7 (1990)
Hong MC, Chang JC, Wu ML, Chang MC, Biochem. Genet, 36, 407 (1998)
Riesenberg D, Schulz V, Knorre WA, Pohl HD, Korz D, Sanders EA, Ross A, Decker WD, J. Biotechnol., 20, 17 (1991)
Koyama Y, Kitao S, Yamamoto-Otake H, Suzuki M, Nakano E, Agric. Biol. Chem., 54, 1453 (1990)
Siegel R, Ryu DDY, Biotechnol. Bioeng., 27, 28 (1985)
Yu HM, Shi Y, Sun XD, Luo H, Shen ZG, J. Biosci. Bioeng., 96(2), 179 (2003)