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Glycoconj J ; 28(8-9): 563-71, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22020441

RESUMEN

A gene for processing α-glucosidase I from a filamentous fungus, Aspergillus brasiliensis (formerly called Aspergillus niger) ATCC 9642 was cloned and fused to a glutathione S-transferase tag. The active construct with the highest production level was a truncation mutant deleting the first 16 residues of the hydrophobic N-terminal domain. This fusion enzyme hydrolyzed pyridylaminated (PA-) oligosaccharides Glc(3)Man(9)GlcNAc(2)-PA and Glc(3)Man(4)-PA and the products were identified as Glc(2)Man(9)GlcNAc(2)-PA and Glc(2)Man(4)-PA, respectively. Saturation curves were obtained for both Glc(3)Man(9)GlcNAc(2)-PA and Glc(3)Man(4)-PA, and the K (m) values for both substrates were estimated in the micromolar range. When 1 µM Glc(3)Man(4)-PA was used as a substrate, the inhibitors kojibiose and 1-deoxynojirimycin had similar effects on the enzyme; at 20 µM concentration, both inhibitors reduced activity by 50%.


Asunto(s)
Aspergillus/enzimología , alfa-Glucosidasas/metabolismo , Acetilglucosamina/metabolismo , Secuencia de Aminoácidos , Aspergillus/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Pruebas de Enzimas , Inhibidores Enzimáticos/farmacología , Vectores Genéticos/genética , Inhibidores de Glicósido Hidrolasas , Hidrólisis/efectos de los fármacos , Interacciones Hidrofóbicas e Hidrofílicas/efectos de los fármacos , Cinética , Datos de Secuencia Molecular , Oligosacáridos/química , Oligosacáridos/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , alfa-Glucosidasas/química
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