title: Enzyme stabilization by covalent binding in nanoporous sol-gel glass for nonaqueous biocatalysis.
abstract: A unique nanoporous sol-gel glass possessing a highly ordered porous structure (with a pore size of 153 A in diameter) was examined for use as a support material for enzyme immobilization. A model enzyme, alpha-chymotrypsin, was efficiently bound onto the glass via a bifunctional ligand, trimethoxysilylpropanal, with an active enzyme loading of 0.54 wt%. The glass-bound chymotrypsin exhibited greatly enhanced stability both in aqueous solution and organic solvents. The half-life of the glass-bound alpha-chymotrypsin was >1000-fold higher than that of the native enzyme, as measured either in aqueous buffer or anhydrous methanol. The enhanced stability in methanol, which excludes the possibility of enzyme autolysis, particularly reflected that the covalent binding provides effective protection against enzyme inactivation caused by structural denaturation. In addition, the activity of the immobilized alpha-chymotrypsin was also much higher than that of the native enzyme in various organic solvents. From these results, it appears that the glass-enzyme complex developed in the present work can be used as a high-performance biocatalyst for various chemical processing applications, particularly in organic media. Published by John Wiley & Sons
authors.affiliation: Department of Chemical Engineering, University of Akron, Akron, Ohio 44325, USA.
authors.forename: P
authors.initials: P
authors.lastname: Wang
authors.forename: S
authors.initials: S
authors.lastname: Dai
authors.forename: S D
authors.initials: SD
authors.lastname: Waezsada
authors.forename: A Y
authors.initials: AY
authors.lastname: Tsao
authors.forename: B H
authors.initials: BH
authors.lastname: Davison
mesh_terms: D002918:Chymotrypsin / Q000737:chemistry* / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D005898:Glass; D012822:Silicon Dioxide; D012997:Solvents; D013499:Surface Properties; D013696:Temperature
chemical_list: D004800:Enzymes, Immobilized; D012997:Solvents; D012822:Silicon Dioxide; D002918:Chymotrypsin; C117608:alpha-chymotrypsin