Identification of essential active-site residues in the cyanogenic beta-glucosidase (linamarase) from cassava (Manihot esculenta Crantz) by site-directed mutagenesis

• Published in: Biochemical journal Vol. 353; no. Pt 2; pp. 199 - 205
• Main Authors: Keresztessy, Z, Brown, K, Dunn, M A, Hughes, M A
• Format: Journal Article
• Language: English
• Published: England 15.01.2001
• Subjects: beta-Glucosidase - biosynthesis; beta-Glucosidase - chemistry; beta-Glucosidase - genetics; Binding Sites; Kinetics; Manihot - enzymology; Manihot - genetics; Models, Molecular; Mutagenesis, Site-Directed; Mutation; Nitriles - chemistry; Recombinant Proteins - biosynthesis; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism

The coding sequence of the mature cyanogenic beta-glucosidase (beta-glucoside glucohydrolase, EC 3.2.1.21; linamarase) was cloned into the vector pYX243 modified to contain the SUC2 yeast secretion signal sequence and expressed in Saccharomyces cerevisiae. The recombinant enzyme is active, glycosyla...