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.
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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
- enzyme activation
- non-aqueous enzymology
- serine protease