Construction and Characterization of Mutant Glucoamylases from the YeastSaccharomycopsis fibuligera

(+)‐Lentiginosine, a Potent and Selective Inhibitor of Amyloglucosidase: Synthetic Efforts and Disputes on Its Absolute Configuration

“…The strongest hydrogen bonds in the X‐ray structure for deoxynojirimycin ( 2 ) at its primary binding site are between O6, O3 and O4 of 2 and Asp55, Arg54 and Leu177. It is worth noting that Asp55 and Arg54 are supposed to be the key residues for the enzyme because mutation experiments led to a complete loss of enzymatic activity 75. Moreover, a crystallographic water molecule (WAT3, Figure 1 a) is directly oriented towards the C1 anomeric carbon atom of the inhibitor.…”

Protein engineering of amylases

“…Random insertion of a BamHI linker DNA was made in the gene of Saccharomycopsisfibuligera glucoamylase [56]. Six out of seven mutants obtained in four regions that are highly conserved among glucoamylases were inactive, while eight out of eleven made outside these regions had activity similar to the wild-type enzyme.…”

Structure/Function Relationships in Starch-Hydrolases and Related Enzymes.

“…Glucoamylases GA from A. awamori52-54) (identical to A. niger GA) and Saccharomycopsis fibuligera55) were mutated either by single amino acid replacement of important residues identified biochemically10) 17) or by sequence alignment, 14,36,55) or by random insertions of a BamHI linker DNA.55)…”