Skip to main content
SpringerLink
Log in

Enhancement of enzymatic catalysis of Bacillus amyloliquefaciens α-amylase by nonionic surfactant micelles

  • Published:
Journal of Surfactants and Detergents

Abstract

Some enzymes are considerably more stable when formulated with nonionic surfactants than when formulated with anionic surfactants. The effect of a nonionic surfatant, polyoxyethylene mono-N-dodecyl ether (Brij 35; number of units of ethylene oxide moieties, 23), on the kinetic behavior of hydrolysis of amylopectin with Bacillus amyloliquefaciens α-amylase (BAA) was studied at a temperature of 25°C and a pH of 7.0. The hydrolytic rate was accelerated by the addition of the nonionic surfactant above its critical micelle concentration. Lineweaver-Burk plots for the enzymatic hydrolysis in the absence and presence of the nonionic surfactant at 0.5 to 2.5% (wt/vol) had linear relationships, and the kinetic parameters, K m and k cat were obtained. The value of k cat was increased with an increased concentration of Brij 35, whereas the K m value was approximately constant. Therefore, the increase in k cat contributed to the acceleration of the apparent hydrolytic rate. The interaction of amylopectin with the surfactant was examined by a surface tension measurement, and the result confirmed the corresponding binding between the substrate and the surfactant. A fluorescence analysis due to tryptophan in BAA suggested that BAA bound to the nonionic micelles. The increase in k cat suggested that hydrolytic catalysis at the micellar pseudophase was more efficient than that at the aqueous pseudophase. The enhancement of the catalytic rate contributed to the effective removal of food stains containing starch when BAA was added with Brij 35 in a laundry detergent washing test.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BAA:

α-amylase from Bacillus amyloliquefaciens

Brij 35:

polyoxyethylene mono-N-dodecyl ether

CMC:

critical micelle concentration

ES complex:

enzyme-substrate complex

References

  1. Kravetz, L., and K.F. Guin, Effect of Surfactant Structure on Stability of Enzymes Formulated into Laundry Liquids, J. Am. Oil Chem. Soc. 62:943 (1985).

    CAS  Google Scholar 

  2. Schomaecker, R., B.H. Robinson, and P.D.I. Fletcher, Interaction of Enzymes with Surfactants in Aqueous Solution and in Water-in-Oil Microemulsions, J. Chem. Soc. Faraday Trans. 1 84:4203 (1988).

    Article  CAS  Google Scholar 

  3. Nakahara, H., S. Okada, K. Mochida, H. Ohmori, and M. Masui, Kinetic Studies on Pancreatic Lipase Activity in Micellar Systems, Chem. Pharm. Bull. 31:4213 (1983).

    CAS  Google Scholar 

  4. Tanaka, A., and E. Hoshino, Thermodynamic and Activation Parameters for the Hydrolysis of Amylose with Bacillus α-Amylases in a Diluted Anionic Surfactant Solution, J. Biosci. Bioeng. 93:485 (2002).

    CAS  Google Scholar 

  5. Montserret, R., M.J. McLeish, A. Böckmann, C. Geourjon, and F. Penin, Involvement of Electrostatic Interactions in the Mechanism of Peptide Folding Induced by Sodium Dodecyl Sulfate Binding, Biochemistry 39:8362 (2000).

    Article  CAS  Google Scholar 

  6. Hagihara, Y., D.-P. Hong, M. Hoshino, K. Enjyoji, H. Kato, and Y. Goto, Aggregation of β2-Glycoprotein I Induced by Sodium Lauryl Sulfate and Lysophospholopids, Biochemistry 41:1020 (2002).

    Article  CAS  Google Scholar 

  7. Russell, G.L., and L.N. Britton, Use of Certain Alcohol Ethoxylates to Maintain Protease Stability in the Presence of Anionic Surfactants, J. Surfact. Deterg. 5:5 (2002).

    CAS  Google Scholar 

  8. Martinek, K., A.V. Levashov, N. Klyachko, Y.L. Khmelnitski, and I.V. Berezin, Micellar Enzymology, Eur. J. Biochem. 155:453 (1986).

    Article  CAS  Google Scholar 

  9. Robyt, J.F., and W.J. Whelan, The α-Amylases, in Starch and Its Derivatives, edited by J.A. Radley, 4th edn., Chapman & Hall, London, 1968, p. 430.

    Google Scholar 

  10. Takagi, T., H. Toda, and T. Isemura, Bacterial and Mold Amylases, in The Enzymes, edited by P.D. Boyer, Academic Press, New York, 1971, p. 235.

    Google Scholar 

  11. Fogarty, W.M., and C.T. Kelly, Amylases, Amyloglucosidases and Related Glucanases, in Microbial Enzymes and Bioconversions, edited by A.H. Rose, Academic Press, London, 1980, p. 115.

    Google Scholar 

  12. Kindle, K.L., Characteristics and Production of Thermostable α-Amylase, Appl. Biochem. Biotechnol. 8:153 (1983).

    CAS  Google Scholar 

  13. Kulp, K., Carbohydrases, in Enzymes in Food Processing, edited by G. Reed, 2nd edn., Academic Press, New York, 1975, p. 53.

    Google Scholar 

  14. Reichelt, J.R. Industrial Applications, in Industrial Enzymology, edited by T. Godfrey, and J.R. Reichelt, Nature Press, New York, 1983, p. 375.

    Google Scholar 

  15. Takkinen, K., R.F. Pettersson, N. Kalkkinen, I. Palva, H. Soderlund, and L. Kaariainen, Amino Acid Sequence of α-Amylase from Bacillus amyloliquefaciens Deduced from the Nucleotide Sequence of the Cloned Gene, J. Biol. Chem. 258:1007 (1983).

    CAS  Google Scholar 

  16. Hiromi, K., Interpretation of Dependency of Rate Parameters on the Degree of Polymerization of Substrate in Enzyme-Catalyzed Reactions. Evaluation of Subsite Affinities of Exoenzyme, Biochem. Biophys. Res. Commun. 40:1 (1970).

    Article  CAS  Google Scholar 

  17. Thoma, J.A., C. Brothers, and J. Spradlin, Subsite Mapping of Enzymes. Studies on Bacillus subtilis Amylase, Biochemistry 9:1768 (1970).

    Article  CAS  Google Scholar 

  18. Fitter, J., R. Herrmann, N.A. Dencher, A. Blume, and T. Hauss, Activity and Stability of a Thermostable α-Amylase Compared to Its Mesophilic Homologue: Mechanisms of Thermal Adaptation, Biochemistry 40:10723 (2001).

    Article  CAS  Google Scholar 

  19. MacGregor, E.A., α-Amylase Structure and Activity, J. Protein Chem. 7:399 (1988).

    Article  CAS  Google Scholar 

  20. Tanaka, A., and E. Hoshino, Study on the Substrate Specificity of α-Amylases That Contribute to Soil Removal in Detergent, J. Surfact. Deterg. 2:193 (1999).

    CAS  Google Scholar 

  21. Viparelli, P., F. Alfani, and M. Cantarella, Models for Enzyme Superactivity in Aqueous Solutions of Surfactants, Biochem. J. 344:765 (1999).

    Article  CAS  Google Scholar 

  22. Svensson, E., M. Gudmundsson, and A.-C. Eliasson, Binding of Sodium Dodecylsulphate to Starch Polysaccharides Quantified by Surface Tension Measurements, Colloids Surf. B 6:227 (1996).

    Article  CAS  Google Scholar 

  23. wulff, G., and S. Kubik, Circular Dichroism and Ultraviolet Spectroscopy of Complexes of Amylose, Carbohydr. Res. 237:1 (1992).

    Article  CAS  Google Scholar 

  24. Hui, Y., J.C. Russell, and D.G. Whitten, Host-Guest Interactions in Amylose Inclusion Complexes, J. Am. Chem. Soc. 105: 1374 (1983).

    Article  CAS  Google Scholar 

  25. Yamamoto, M., S. Harada, T. Nakatsuka, and T. Sano, Hydrodynamic Characterization of Amylose-SDS Complex by Viscosity Measurement, Bull. Chem. Soc. Jpn. 61:1471 (1988).

    Article  CAS  Google Scholar 

  26. Burstein, E.A., N.S. Vedenkina, and M.N. Ivkova, Fluorescence and the Location of Tryptophan Residues in Protein Molecules, Photochem. Photobiol. 18:263 (1973).

    CAS  Google Scholar 

  27. Suzuki, A., T. Yamane, Y. Ito, T. Nishio, H. Fujiwara, and T. Ashida, Crystallization and Preliminary Crystallographic Study of Bacterial α-Amylases, J. Biochem. 108:379 (1990).

    CAS  Google Scholar 

  28. Bernfeld, P., Amylases, α and β, Methods Enzymol 1:149 (1955).

    Google Scholar 

  29. Padday, J.F., The Measurement of Surface Tension, in Surface and Colloid Science Vol. 1, edited by E. Matijevic, Wiley-Interscience, New York, 1969, p. 101.

    Google Scholar 

  30. Fendler, J.H., and E.J. Fendler, Principles of Micellar Catalysis in Aqueous Solutions, in Catalysis in Micellar and Macromolecular Systems, Academic Press, London, 1975, p. 86.

    Google Scholar 

  31. Torgerson, E.M., L.C. Brewer, and J.A. Thoma, Subsite Mapping of Enzymes. Use of Subsite Map to Simulate Complete Time Course of Hydrolysis of a Polymeric Substrate, Arch. Biochem. Biophys. 196:13 (1979).

    Article  CAS  Google Scholar 

  32. Dua, R.D., and S. Kochhar, Active Site Studies on Bacillus amyloliquefaciens α-Amylase, Mol. Cell. Biochem. 66:13 (1985)

    Article  CAS  Google Scholar 

  33. Ooshima, H., M. Sakata, and Y. Harano, Enhancement of Enzymatic Hydrolysis of Cellulose by Surfactant, Biotech. Bioeng. 28:1727 (1986).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Tokyo Research Laboratories, Kao Corporation, 31-8501, Tokyo, Japan

    Eiichi Hoshino

  2. Household Products Research Laboratories, Kao Corp., 1334 Minato, Wakayama-shi, Wakayama, 640-8580, Japan

    Atsushi Tanaka

Authors
  1. Eiichi Hoshino
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Atsushi Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Atsushi Tanaka.

About this article

Cite this article

Hoshino, E., Tanaka, A. Enhancement of enzymatic catalysis of Bacillus amyloliquefaciens α-amylase by nonionic surfactant micelles. J Surfact Deterg 6, 299–303 (2003). https://doi.org/10.1007/s11743-003-0273-2

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11743-003-0273-2

Key Words

Search

Navigation