Mutations of Trp275 and Trp397 altered the binding selectivity of Vibrio carchariae chitinase A

• Published in: Biochimica et biophysica acta Vol. 1770; no. 8; pp. 1151 - 1160
• Main Authors: Suginta, Wipa, Songsiriritthigul, Chomphunuch, Kobdaj, Archara, Opassiri, Rodjana, Svasti, Jisnuson
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.08.2007
• Subjects: Active-site mutation; Binding Sites; Biological Assay; Chitin - metabolism; Chitin hydrolysis; Chitinase A; Chitinases - chemistry; Chitinases - metabolism; Chitooligosaccharide; Chromatography, Thin Layer; Circular Dichroism; Escherichia coli - genetics; Hydrolysis; Kinetics; Models, Molecular; Mutagenesis, Site-Directed; Point Mutation; Protein Binding; Protein Conformation; Specific hydrolyzing activity; Substrate Specificity; Thin layer chromatography; Tryptophan - chemistry; Tryptophan - genetics; Vibrio - enzymology; Vibrio carchariae; Chitinase A; Gn; TLC; DMAB; Thin layer chromatography; Vibrio carchariae; PMSF; Chitin hydrolysis; Chitooligosaccharide; IPTG; Specific hydrolyzing activity; Active-site mutation; pNP-(GlcNAc) 2

Point mutations of the active-site residues Trp168, Tyr171, Trp275, Trp397, Trp570 and Asp392 were introduced to Vibrio carchariae chitinase A. The modeled 3D structure of the enzyme illustrated that these residues fully occupied the substrate binding cleft and it was found that their mutation great...