Korean Journal of Chemical Engineering, Vol.25, No.5, 1110-1114, September, 2008
A fermentation strategy for production of recombinant protein subjected to plasmid instability
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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.
- 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)