Biotechnology Letters

, Volume 30, Issue 4, pp 701–706

Impact of surfactants on solubilization and activity of the carotenoid cleavage dioxygenase, AtCCD1, in an aqueous micellar reaction system

Original Research Paper


The effect of various surfactants on both the solubilization of the carotenoid cleavage dioxygenase, AtCCD1, from cell lysates and the enzymatic activity in an aqueous micellar system was investigated. Solubilization with sodium cholate more than doubled the specific activity. Lag phases were observed when Tween surfactants were used for substrate delivery and were dependent on the surfactant and enzyme modification. In contrast to His6- and GST-tagged AtCCD1, unmodified AtCCD1 showed a 45% increased maximum rate in the Tween 20 system compared to Triton X-100 based reference system. The results emphasize the importance of engineering the interface for the in vitro application of this enzyme family.


Carotenoid cleavage dioxygenase Micelles Non-ionic surfactants Protein solubilization Surfactants 


  1. Bhattacharya SC, Palepu RM (2004) Micellization of nonionic surfactants: tweens––a review. J Surface Sci Technol 20:159–177Google Scholar
  2. Carman GM, Deems RA, Dennis EA (1995) Lipid signaling enzymes and surface dilution kinetics. J Biol Chem 270:18711–18714PubMedCrossRefGoogle Scholar
  3. Dwars T, Paetzold E, Oehme G (2005) Reactions in micellar systems. Angew Chem Int Ed Engl 44:7174–7199PubMedCrossRefGoogle Scholar
  4. Lim WH, Lawrence MJ (2004) Influence of surfactant and lipid chain length on the solubilisation of phosphatidylcholine vesicles by micelles comprised of polyoxyethylene sorbitan monoesters. Colloids Surf A 250:449–457CrossRefGoogle Scholar
  5. Marasco EK, Vay K, Schmidt-Dannert C (2006) Identification of carotenoid cleavage dioxygenases from Nostoc sp. PCC 7120 with different cleavage activities. J Biol Chem 281:31583–31593PubMedCrossRefGoogle Scholar
  6. Molina-Bolivar JA, Aguiar J, Peula-Garcia JM et al (2002) Photophysical and light scattering studies on the aggregation behaviour of Triton X-100 in formamide-water mixed solvents. Mol Phys 100:3259–3269CrossRefGoogle Scholar
  7. Patist A, Kanicky JR, Shukla PK et al (2002) Importance of micellar kinetics in relation to technological processes. J Colloid Interface Sci 245:1–15PubMedCrossRefGoogle Scholar
  8. Sambrook J, Russell DW (2001) Molecular cloning––a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  9. Schilling M, Patett F, Schwab W et al (2007) Influence of solubility-enhancing fusion proteins and organic solvents on the in vitro biocatalytic performance of the carotenoid cleavage dioxygenase AtCCD1 in a micellar reaction system. Appl Microbiol Biotechnol 75:829–836PubMedCrossRefGoogle Scholar
  10. Schmidt H, Kurtzer R, Eisenreich W et al (2006) The carotenase AtCCD1 from Arabidopsis thaliana is a dioxygenase. J Biol Chem 281:9845–9851PubMedCrossRefGoogle Scholar
  11. Schwartz SH, Qin X, Zeevaart JA (2001) Characterization of a novel carotenoid cleavage dioxygenase from plants. J Biol Chem 276:25208–25211PubMedCrossRefGoogle Scholar
  12. Viparelli P, Alfani F et al (1999) Models for enzyme superactivity in aqueous solutions of surfactants. Biochem J 344:765–773PubMedCrossRefGoogle Scholar
  13. Wang ZX, Killilea SD, Srivastava DK (1993) Kinetic evaluation of substrate-dependent origin of the lag phase in soybean lipoxygenase-1 catalyzed reactions. Biochemistry 32:1500–1509PubMedCrossRefGoogle Scholar
  14. Yamazaki K, Imai M, Suzuki I (2004) Soybean lecithin hydrolysis using hog pancreas phospholipase A2 influenced by the hydrophobic character of W/O microemulsion systems. Biochem Eng J 19:171CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • M. Schilling
    • 1
  • F. Haetzelt
    • 1
  • W. Schwab
    • 2
  • J. Schrader
    • 1
  1. 1.Biochemical EngineeringDECHEMA e.V., Karl-Winnacker-InstitutFrankfurt/MainGermany
  2. 2.Biomolecular Food TechnologyTU MünchenFreisingGermany

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