DNA Plasmid Production in Different Host Strains of Escherichia Coli

Overview

Journal J Ind Microbiol Biotechnol

Publisher Oxford University Press

Specialty Biotechnology

Date 2009 Jan 13

PMID 19137335

Citations 10

Authors

Adam Singer

Mark A Eiteman

Elliot Altman

Affiliations

Soon will be listed here.

Abstract

We compared plasmid DNA production in 13 strains of Escherichia coli in shake flasks using media containing glucose or glycerol. DNA yield from either carbon source showed small correlation with maximum growth rate. Three strains, SCS1-L, BL21 and MC4100, were selected for a controlled exponential fed-batch process at a growth rate of 0.14 h(-1) to an optical density of about 70, followed by a four-hour heat treatment. Prior to heat treatment, SCS1-L generated 15.4 mg DNA/g, BL21 generated 11.0 mg DNA/g and MC4100 generated 7.9 mg DNA/g, while after heat treatment the strains attained DNA yields, respectively, of 18.0, 15.0 and 6.8 mg/g. The strains also varied in their percentage of supercoiled DNA after heat treatment, with SCS1-L averaging 66% supercoiled, BL21 17% and MC4100 40%. We further investigated the two strains that yielded the highest percentage of supercoiled DNA (SCS1-L and MC4100) at a higher growth rate of 0.28 h(-1). At this condition, a slightly lower DNA yield was generated faster, and the percentage of supercoiled DNA increased. Heat treatment improved DNA yield, and surprisingly did so to a greater extent at the higher growth rate. As a consequence of these factors, higher growth rates might be advantageous for DNA production.

Citing Articles

Enabling mRNA Therapeutics: Current Landscape and Challenges in Manufacturing.

Youssef M, Hitti C, Puppin Chaves Fulber J, Kamen A Biomolecules. 2023; 13(10).

PMID: 37892179 PMC: 10604719. DOI: 10.3390/biom13101497.

To Plate or to Simply Unfreeze, That Is the Question for Optimal Plasmid Extraction.

Thean R, Ong D, Heng Z, Gan S, Yeo J J Biomol Tech. 2021; 32(2):57-62.

PMID: 34121935 PMC: 8174125. DOI: 10.7171/jbt.20-3203-001.

High Kanamycin Concentration as Another Stress Factor Additional to Temperature to Increase pDNA Production in DH5α Batch and Fed-Batch Cultures.

Grijalva-Hernandez F, Vega-Estrada J, Escobar-Rosales M, Ortega-Lopez J, Aguilar-Lopez R, Lara A Microorganisms. 2019; 7(12).

PMID: 31861108 PMC: 6955755. DOI: 10.3390/microorganisms7120711.

Application of Plasmid Engineering to Enhance Yield and Quality of Plasmid for Vaccine and Gene Therapy.

Folarin O, Nesbeth D, Ward J, Keshavarz-Moore E Bioengineering (Basel). 2019; 6(2).

PMID: 31248216 PMC: 6631426. DOI: 10.3390/bioengineering6020054.

Optimization of fermentation conditions for an strain engineered using the response surface method to produce a novel therapeutic DNA vaccine for rheumatoid arthritis.

Long J, Zhao X, Liang F, Liu N, Sun Y, Xi Y J Biol Eng. 2018; 12:22.

PMID: 30337953 PMC: 6180442. DOI: 10.1186/s13036-018-0110-y.

References

Miki T, Yasukochi T, Nagatani H, Furuno M, Orita T, Yamada H . Construction of a plasmid vector for the regulatable high level expression of eukaryotic genes in Escherichia coli: an application to overproduction of chicken lysozyme. Protein Eng. 1987; 1(4):327-32. DOI: 10.1093/protein/1.4.327. View

Xu Z, Shen W, Chen H, Cen P . Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy. J Zhejiang Univ Sci B. 2005; 6(5):396-400. PMC: 1389757. DOI: 10.1631/jzus.2005.B0396. View

Arnold C, McElhanon J, Lee A, Leonhart R, Siegele D . Global analysis of Escherichia coli gene expression during the acetate-induced acid tolerance response. J Bacteriol. 2001; 183(7):2178-86. PMC: 95122. DOI: 10.1128/JB.183.7.2178-2186.2001. View

Shamlou P . Scaleable processes for the manufacture of therapeutic quantities of plasmid DNA. Biotechnol Appl Biochem. 2003; 37(Pt 3):207-18. DOI: 10.1042/BA20030011. View

Wang Z, Yuan Z, Hengge U . Processing of plasmid DNA with ColE1-like replication origin. Plasmid. 2004; 51(3):149-61. DOI: 10.1016/j.plasmid.2003.12.002. View

Gilbert S, Moorthy V, Andrews L, Pathan A, McConkey S, Vuola J . Synergistic DNA-MVA prime-boost vaccination regimes for malaria and tuberculosis. Vaccine. 2005; 24(21):4554-61. DOI: 10.1016/j.vaccine.2005.08.048. View

Listner K, Bentley L, Okonkowski J, Kistler C, Wnek R, Caparoni A . Development of a highly productive and scalable plasmid DNA production platform. Biotechnol Prog. 2006; 22(5):1335-45. DOI: 10.1021/bp060046h. View

Yau S, Keshavarz-Moore E, Ward J . Host strain influences on supercoiled plasmid DNA production in Escherichia coli: Implications for efficient design of large-scale processes. Biotechnol Bioeng. 2008; 101(3):529-44. DOI: 10.1002/bit.21915. View

Fitzwater T, Zhang X, Elble R, Polisky B . Conditional high copy number ColE1 mutants: resistance to RNA1 inhibition in vivo and in vitro. EMBO J. 1988; 7(10):3289-97. PMC: 454759. DOI: 10.1002/j.1460-2075.1988.tb03196.x. View

10.

Bentley W, Mirjalili N, Andersen D, Davis R, Kompala D . Plasmid-encoded protein: the principal factor in the "metabolic burden" associated with recombinant bacteria. Biotechnol Bioeng. 1990; 35(7):668-81. DOI: 10.1002/bit.260350704. View

11.

Siegel R, Ryu D . Kinetic study of instability of recombinant plasmid pPLc23trpAl in E. coli using two-stage continuous culture system. Biotechnol Bioeng. 1985; 27(1):28-33. DOI: 10.1002/bit.260270105. View

12.

Seo J, Bailey J . Effects of recombinant plasmid content on growth properties and cloned gene product formation in Escherichia coli. Biotechnol Bioeng. 1985; 27(12):1668-74. DOI: 10.1002/bit.260271207. View

13.

Wong E, Muesing M, Polisky B . Temperature-sensitive copy number mutants of CoIE1 are located in an untranslated region of the plasmid genome. Proc Natl Acad Sci U S A. 1982; 79(11):3570-4. PMC: 346463. DOI: 10.1073/pnas.79.11.3570. View

14.

Okonkowski J, Kizer-Bentley L, Listner K, Robinson D, Chartrain M . Development of a robust, versatile, and scalable inoculum train for the production of a DNA vaccine. Biotechnol Prog. 2005; 21(4):1038-47. DOI: 10.1021/bp040041p. View

15.

Nichols W, Ledwith B, Manam S, Troilo P . Potential DNA vaccine integration into host cell genome. Ann N Y Acad Sci. 1995; 772:30-9. DOI: 10.1111/j.1749-6632.1995.tb44729.x. View

16.

Eiteman M, Altman E . Overcoming acetate in Escherichia coli recombinant protein fermentations. Trends Biotechnol. 2006; 24(11):530-6. DOI: 10.1016/j.tibtech.2006.09.001. View

17.

Donnelly J, Berry K, Ulmer J . Technical and regulatory hurdles for DNA vaccines. Int J Parasitol. 2003; 33(5-6):457-67. DOI: 10.1016/s0020-7519(03)00056-0. View

18.

Weintraub H, Cheng P, Conrad K . Expression of transfected DNA depends on DNA topology. Cell. 1986; 46(1):115-22. DOI: 10.1016/0092-8674(86)90865-2. View

19.

Lahijani R, Hulley G, Soriano G, Horn N, Marquet M . High-yield production of pBR322-derived plasmids intended for human gene therapy by employing a temperature-controllable point mutation. Hum Gene Ther. 1996; 7(16):1971-80. DOI: 10.1089/hum.1996.7.16-1971. View

20.

Engberg B, Nordstrom K . Replication of R-factor R1 in Scherichia coli K-12 at different growth rates. J Bacteriol. 1975; 123(1):179-86. PMC: 235705. DOI: 10.1128/jb.123.1.179-186.1975. View