Applied Microbiology and Biotechnology

, Volume 83, Issue 3, pp 589–596 | Cite as

Use of cyclodextrin and its derivatives for increased transformation efficiency of competent bacterial cells

  • Finn Lillelund AachmannEmail author
  • Trond Erik Vee Aune


Methodologies for introduction of DNA into cells are essential in molecular genetics and vital for applications such as genetic engineering and gene therapy. The use of cyclodextrins (CyDs) for increased efficiency of introducing DNA into eukaryotic cells (transfection) has been reported, but CyDs’ effect on the introduction of DNA into bacterial cells (transformation) is unknown. Here, we have investigated the potential of using CyDs in the transformation of chemically competent in-house, commercially available, and, on non-competent bacterial cells, with plasmid DNA of two different sizes. Possible interactions between CyDs and DNA were studied with nuclear magnetic resonance (NMR) spectroscopy. The presence of CyDs resulted in an up to fourfold increment of the transformation rate for in-house cells, with β-CyD and derivates giving the strongest effect. For commercial cells and transformation with megaplasmids, a more moderate effect around 1.4-fold was obtained. However, CyDs have little or no effect on DNA uptake by noncompetent cells. Results obtained from NMR spectroscopy show no interactions between CyDs and DNA-like molecules, which indicated that the CyDs’ effect is related to the bacterial cell wall.


Cyclodextrins DNA Plasmid Bacteria Competent cell Transformation 



We thank Technology Transfer Office for financial support. Furthermore, we thank Svein Valla and Kim L. Larsen for proofreading the manuscript and for useful discussions and comments.


  1. Bar R (1989) Cyclodextrin-aided microbial transformation of aromatic-aldehydes by Saccharomyces-Cerevisiae. Appl Microbiol Biot 31:25–28CrossRefGoogle Scholar
  2. Bar R, Ulitzur S (1994) Bacterial toxicity of cyclodextrins—luminous Escherichia coli as a model. Appl Microbiol Biot 41:574–577CrossRefGoogle Scholar
  3. Bartlett DW, Davis ME (2007) Physicochemical and biological characterization of targeted, nucleic acid-containing nanoparticles. Bioconjug Chem 18:456–468CrossRefGoogle Scholar
  4. Chadha R, Kapoor VK et al (2008) Drug carrier systems for anticancer agents: A review. J Sci Ind Res India 67:185–197Google Scholar
  5. Cohen SN, Chang AC et al (1972) Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci USA 69:2110–2114CrossRefGoogle Scholar
  6. Cohen SN, Chang AC et al (1973) Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci USA 70:3240–3244CrossRefGoogle Scholar
  7. Connors KA (1997) The Stability of Cyclodextrin Complexes in Solution. Chem Rev 97:1325–1357CrossRefGoogle Scholar
  8. Easton CJ, Lincoln SF (1999) Modified Cyclodextrins. Imperial College Press, LondonGoogle Scholar
  9. Galant C, Amiel C et al (2005) Ternary complex formation in aqueous solution between a beta-cyclodextrin polymer, a cationic surfactant and DNA. Macromol Biosci 5:1057–1065CrossRefGoogle Scholar
  10. Graham DR, Chertova E et al (2003) Cholesterol depletion of human immunodeficiency virus type 1 and simian immunodeficiency virus with beta-cyclodextrin inactivates and permeabilizes the virions: evidence for virion-associated lipid rafts. J Virol 77:8237–8248CrossRefGoogle Scholar
  11. Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580CrossRefGoogle Scholar
  12. Hanahan D, Jessee J et al (1991) Plasmid transformation of Escherichia coli and other bacteria. Methods Enzymol 204:63–113CrossRefGoogle Scholar
  13. Ilangumaran S, Hoessli DC (1998) Effects of cholesterol depletion by cyclodextrin on the sphingolipid microdomains of the plasma membrane. Biochem J 335:433–440Google Scholar
  14. Irie T, Uekama K (1999) Cyclodextrn in pepide and Prortein delivery. Adv Drug Deliv Rev 36:101–123CrossRefGoogle Scholar
  15. Irie T, Fukunaga K et al (1992a) Hydroxypropylcyclodextrins in parenteral use. I: Lipid dissolution and effects on lipid transfers in vitro. J Pharm Sci 81:521–523CrossRefGoogle Scholar
  16. Irie T, Wakamatsu K et al (1992b) Enhancing effects of cyclodextrins on nasal absorption of insulin in rats. Int J Pharm 84:129CrossRefGoogle Scholar
  17. Khanna KV, Whaley KJ et al (2002) Vaginal transmission of cell-associated HIV-1 in the mouse is blocked by a topical, membrane-modifying agent. J Clin Invest 109:205–211Google Scholar
  18. Kilsdonk EPC, Yancey PG et al (1995) Cellular cholesterol efflux mediated by cyclodextrins. J Biol Chem 270:17250–17256CrossRefGoogle Scholar
  19. Larsen KL (2002) Large cyclodextrins. J Inclusion Phenom Macro 43:1–13CrossRefGoogle Scholar
  20. Loftsson T, Vogensen SB et al (2007) Effects of cyclodextrins on drug delivery through biological membranes. J Pharm Sci 96:2532–2546CrossRefGoogle Scholar
  21. Mandel M, Higa A (1970) Calcium-dependent bacteriophage DNA infection. J Mol Biol 53:159–162CrossRefGoogle Scholar
  22. Niyogi K, Hildreth JE (2001) Characterization of new syncytium-inhibiting monoclonal antibodies implicates lipid rafts in human T-cell leukemia virus type 1 syncytium formation. J Virol 75:7351–7361CrossRefGoogle Scholar
  23. Norgard MV, Keem K et al (1978) Factors affecting the transformation of Escherichia coli strain chi1776 by pBR322 plasmid DNA. Gene 3:279–292CrossRefGoogle Scholar
  24. Ohtani Y, Irie T et al (1989) Differential effects of alpha-, beta- and gamma-cyclodextrins on human erythrocytes. Eur J Biochem 186:17–22CrossRefGoogle Scholar
  25. Pack DW, Hoffman AS et al (2005) Design and development of polymers for gene delivery. Nat Rev Drug Discov 4:581–593CrossRefGoogle Scholar
  26. Panja S, Saha S et al (2006) Role of membrane potential on artificial transformation of E. coli with plasmid DNA. J Biotechnol 127:14–20Google Scholar
  27. Redenti E, Pietra C et al (2001) Cyclodextrins in oligonucleotide delivery. Adv Drug Deliv Rev 53:235–244CrossRefGoogle Scholar
  28. Robyt JF (1998) Essentials of Carbohydrate Chemistry. Springer, BerlinGoogle Scholar
  29. Rodal SK, Skretting G et al (1999) Extraction of cholesterol with methyl-beta-cyclodextrin perturbs formation of clathrin-coated endocytic vesicles. Mole Biol Cell 10:961–974Google Scholar
  30. Rothblat GH, de la Llera-Moya M et al (1999) Cell cholesterol efflux: integration of old and new observations provides new insights. J Lipid Res 40:781–796Google Scholar
  31. Sambrook J, Russell D (2001) In Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  32. Solomon JM, Grossman AD (1996) Who’s competent and when: regulation of natural genetic competence in bacteria. Trends Genet 12:150–155CrossRefGoogle Scholar
  33. Spies MA, Schowen RL (2002) The trapping of a spontaneously “flipped-out” base from double helical nucleic acids by host-guest complexation with beta-cyclodextrin: The intrinsic base-flipping rate constant for DNA and RNA. J Am Chem Soc 124:14049–14053CrossRefGoogle Scholar
  34. Szejtli J (1998) Introduction and General Overview of Cyclodextrin Chemistry. Chem Rev 98:1743–1753CrossRefGoogle Scholar
  35. Tsutsumi T, Hirayama F et al (2007a) Evaluation of polyamidoamine dendrimer/alpha-cyclodextrin conjugate (generation 3, G3) as a novel carrier for small interfering RNA (siRNA). J Control Release 119:349–359CrossRefGoogle Scholar
  36. Tsutsumi T, Hirayama F et al (2007b) Potential use of polyamidoamine dendrimer/alpha-cyclodextrin conjugate (generation 3, G3) as a novel carrier for short hairpin RNA-expressing plasmid DNA. J Pharm Sci 97:3022–3034CrossRefGoogle Scholar
  37. Wimmer R, Aachmann FL et al (2002) NMR diffusion as a novel tool for measuring the association constant between cyclodextrin and guest molecules. Carbohydr Res 337:841–849CrossRefGoogle Scholar
  38. Yancey PG, Rodriqueza WV et al (1996) Cellular cholesterol effect mediated by cyclodextrins - Demonstration of kinetic pools and mechanism of efflux. J Biol Chem 271:16026–16034CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Finn Lillelund Aachmann
    • 1
    Email author
  • Trond Erik Vee Aune
    • 1
  1. 1.NOBIPOL, Department of BiotechnologyNorwegian University of Science and TechnologyTrondheimNorway

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