| Title | Cloning, expression, characterization, and nucleophile identification of family 3, Aspergillus niger beta-glucosidase |
| Publication Type | Journal Article |
| Year of Publication | 2000 |
| Authors | Dan, S, Marton, I, Dekel, M, Bravdo, BA, He, SM, Withers, SG, Shoseyov, O |
| Journal | JOURNAL OF BIOLOGICAL CHEMISTRY |
| Volume | 275 |
| Pagination | 4973-4980 |
| Date Published | FEB 18 |
| ISSN | 0021-9258 |
| Abstract | {The beta-glucosidase from Aspergillus niger (CMI. CC 324262) was purified, and an N-terminal sequence and two internal sequences were determined. BglI genomic gene and the cDNA were cloned from a genomic library and by reverse transcriptase-polymerase chain reaction, respectively. The cDNA was successfully expressed in Saccharomyces cerevisiae and Pichia pastoris. Sequence analysis revealed that the gene encodes a 92-kDa enzyme that is a member of glycosidase family 3. H-1-NMR analysis of the reaction catalyzed by this enzyme confirmed that, in common with other family 3 glycosidases, this enzyme hydrolyzes with net retention of anomeric configuration. Accordingly, the enzyme was inactivated by a-deoxy-a-fluoro beta-glucosyl fluoride, with kinetic parameters of k(i) = 4.5 min(-1) |
| DOI | 10.1074/jbc.275.7.4973 |
