Skip to main content
Log in

Activation of serine protease subtilisin Carlsberg in organic solvents: combined effect of methyl-β-cyclodextrin and water

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Methyl β-cyclodextrin (MβCD) increased the activity and enantioselectivity of lyophilized subtilisin suspended in dry THF and acetonitrile in two transesterification model reactions. These beneficial improvements were diminished by the addition of water, in contrast to the observation that water activates subtilisin lyophilized from buffer alone. For example, the initial rate for the S enantiomer in the transesterification of vinylbutyrate with (±)-1-phenylethanol (sec-phenethylalcohol) decreased ca. 4-fold and the enantioselectivity from 59 to 40 when 0.1% (v/v) of water was added to THF.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bell G, Halling PJ, Moore BD, Partridge J, Rees DG (1995) Biocatalyst behavior in low-water systems. Trends Biotechnol. 13: 468–473.

    Google Scholar 

  • Broos J, Sakodinskaya IK, Engbersen JFJ, Verboom W, Reinhoudt DN (1995) Large activation of serine proteases by pretreatment with crown ethers. J. Chem. Soc. Chem. Comm. 255–256.

  • Dabulis K, Klibanov AM (1993) Dramatic enhancement of enzymatic activity in organic solvents by lyoprotectants. Biotechnol. Bioeng. 41: 566–571.

    Google Scholar 

  • Engbersen JFJ, Broos J, Verboom W, Reinhoudt DN (1996) Effects of crown ethers and small amounts of cosolvent on the activity and enantioselectivity of α-chymotrypsin in organic solvents. Pure Appl. Chem. 68: 2171–2178.

    Google Scholar 

  • Greenspan L (1977) Humidity fixed points of binary saturated aqueous solutions. J. Res. Natl. Bureau of Standards — A. Phys. Chem. 81A: 89–96.

    Google Scholar 

  • Griebenow K, Klibanov AM (1996) On protein denaturation in aqueous-organic but not in pure organic solvents. J. Am. Chem Soc. 118: 11695–11700.

    Google Scholar 

  • Griebenow K, Díaz Laureano Y, Santos AM, Montañez Clemente I, Rodríguez L, Vidal M, Barletta G (1999) Improved enzyme activity and enantioselectivity in organic solvents by methyl-β-cyclodextrin. J. Am. Chem. Soc. 121: 8157–8163.

    Google Scholar 

  • Halling PJ (1994) Thermodynamic predictions for biocatalysis in nonconventional media: theory, tests, and recommendations for experimental design and analysis. Enzyme Microb. Technol. 16: 178–206.

    Google Scholar 

  • Klibanov AM (1990) Asymmetric transformations catalyzed by enzymes in organic solvents. Acc. Chem. Res. 23: 114–120.

    Google Scholar 

  • Klibanov AM (1997) Why are enzymes less active in organic solvents than in water? Trends Biotechnol. 15: 97–101.

    Google Scholar 

  • Partridge J, Dennison PR, Moore BD, Halling PJ (1998) Activity and mobility of subtilisin in low water organic media: hydration is more important than solvent dielectric. Biochim. Biophys. Acta 1386: 79–89.

    Google Scholar 

  • Schmitke JL, Wescott CR, Klibanov AM (1996) The mechanistic dissection of the plunge in enzymatic activity upon transition from water to anhydrous solvents. J. Am. Chem. Soc. 118: 3360–3365.

    Google Scholar 

  • van Unen DJ, Engbersen JFJ, Reinhoudt DN (1998) Large acceleration of α-chymotrypsin-catalyzed dipeptide formation by 18-crown-6 in organic solvents. Biotechnol. Bioeng. 59: 553–556.

    Google Scholar 

  • Zaks A, Klibanov AM (1988) Enzymatic catalysis in nonaqueous solvents. J. Biol. Chem. 263: 3194–3201.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Santos, A.M., Montañez Clemente, I., Barletta, G. et al. Activation of serine protease subtilisin Carlsberg in organic solvents: combined effect of methyl-β-cyclodextrin and water. Biotechnology Letters 21, 1113–1118 (1999). https://doi.org/10.1023/A:1005626211015

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005626211015

Navigation