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Literature summary extracted from

  • Sauer, J.; Abou Hachem, M.; Svensson, B.; Jensen, K.J.; Thygesen, M.B.
    Kinetic analysis of inhibition of glucoamylase and active site mutants via chemoselective oxime immobilization of acarbose on SPR chip surfaces (2013), Carbohydr. Res., 375, 21-28.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.2.1.3 analysis method for anchoring native acarbose to gold chip surfaces for surface plasmon resonance studies employing glucoamylase as analyte. The key method is the chemoselective and protecting group-free oxime functionalization of the pseudo-tetrasaccharide-based inhibitor acarbose. At pH 7.0 the association and dissociation rate constants for the acarbose-glucoamylase interaction are 10000 per M and s and 103 per s, respectively, and the conformational change to a tight enzyme-inhibitor complex affects the dissociation rate constant by a factor of 100 per s. The acarbose-presenting surface plason resonance surfaces can be used as a glucoamylase sensor that allows rapid, label-free affinity screening of small carbohydrate-based inhibitors in solution Aspergillus niger

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.3 E180Q active site mutant. For inhibitor acarbose, a rapid binding event is apparently intersected by a slower secondary binding event. Mutant shows a dramatically higher Kd value for acarbose Aspergillus niger
3.2.1.3 R54L active site mutant. For inhibitor acarbose, a rapid binding event is apparently intersected by a slower secondary binding event. Mutant shows a dramatically higher Kd value for acarbose Aspergillus niger
3.2.1.3 Y175F mutation in subsite +3. Mutant displays only minor differences to wild-type in affinities to inhibitors acarbose and an acarbose conjugate Aspergillus niger

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.3 1-deoxynojirimycin
-
 Aspergillus niger
3.2.1.3 acarbose
-
 Aspergillus niger

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.3 Aspergillus niger
-
-
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.3 commercial preparation
-
 Aspergillus niger
-