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%.

N-Linked oligosaccharide, Aspergillus brasiliensis, Processing α-glucosidase I, Pyridylaminated oligosaccharide, Kojibiose

• Compound ID: 18624
  

  a-D-Manp-(1-2)-a-D-Manp-(1-6)-+ | a-D-Manp-(1-2)-a-D-Manp-(1-3)-a-D-Manp-(1-6)-+ | a-D-Glcp-(1-2)-a-D-Glcp-(1-3)-a-D-Glcp-(1-3)-a-D-Manp-(1-2)-a-D-Manp-(1-2)-a-D-Manp-(1-3)-b-D-Manp-(1-4)-b-D-GlcpNAc-(1-4)-b-D-GlcpNAc1N-(1--/pyridylamine/

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  Structure type: oligomer
  Aglycon: pyridylamine
  
   
  
  Aspergillus brasiliensis ATCC 9642
  CSDB ID 43605 (all data & tools)