Enzyme stabilization by covalent binding in nanoporous sol-gel glass for nonaqueous biocatalysis

Ping Wang, Sheng Dai, S. D. Waezsada, Alice Y. Tsao, Brian H. Davison

Research output: Contribution to journalArticlepeer-review

186 Scopus citations

Abstract

A unique nanoporous sol-gel glass possessing a highly ordered porous structure (with a pore size of 153 Å in diameter) was examined for use as a support material for enzyme immobilization. A model enzyme, α-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 α-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 α-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.

Original languageEnglish
Pages (from-to)249-255
Number of pages7
JournalBiotechnology and Bioengineering
Volume74
Issue number3
DOIs
StatePublished - Aug 6 2001

Keywords

  • Enzyme immobilization
  • Nonaqueous biocatalysis
  • Organic solvents
  • Sol-gel silica glass
  • α-chymotrypsin

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