|Title||REGION-DIRECTED MUTAGENESIS OF RESIDUES SURROUNDING THE ACTIVE-SITE NUCLEOPHILE IN BETA-GLUCOSIDASE FROM AGROBACTERIUM-FAECALIS|
|Publication Type||Journal Article|
|Year of Publication||1992|
|Authors||TRIMBUR, DE, WARREN, RAJ, Withers, SG|
|Journal||JOURNAL OF BIOLOGICAL CHEMISTRY|
|Date Published||MAY 25|
The active site nucleophile of the beta-glucosidase of Agrobacterium faecalis has recently been identified by the use of inhibitors. A combination of site-directed and in vitro enzymatic mutagenesis was carried out on the beta-glucosidase to probe the structure of the active site region. Forty-three point mutations were generated at 22 different residues in the region surrounding the active site nucleophile, Glu358. Only five positions were identified which affected enzyme activity indicating that only a few key residues are important to enzyme activity, thus the enzyme can tolerate a number of single residue changes and still function. The importance of Glu358 to enzymatic function has been confirmed and other residues important to enzyme structure or function have been identified.