Enhancing the catalytic efficiency of subtilisin for transesterification by dual bioimprinting.

Bioimprinting is a technique in which an aqueous solution of a protein molecule along with the imprint molecule is dried to remove bulk water. Subtilisin was dissolved in an aqueous buffer with a substrate analog and precipitated with the substrate alcohol to obtain a dually bioimprinted enzyme precipitate. Precipitation was found to be a better bioimprinting technique than freeze drying used so far. The precipitation also enabled simultaneous bioimprinting with the alcohol substrate reinforcing the existing imprint with the substrate analog. The dually bioimprinted subtilisin was found out to be 65-fold more efficient than the freeze dried non-imprinted powder of subtilisin and could transesterify N -acetyl- l -phenylalanine with n -propanol up to 82% in 5 h. [ABSTRACT FROM AUTHOR]