Characterization of the GPI-anchored endo β-1,3-glucanase Eng2 of Aspergillus fumigatus.
A GPI-anchored endo β-1,3-glucanase of Aspergillus fumigatus was characterized. The enzyme encoded by ENG2 (AFUA_2g14360) belongs to the glycoside hydrolase family 16 (GH16). The activity was characterized using a recombinant protein produced by Pichiapastoris. The recombinant enzyme preferentially acts on soluble β-1,3-glucans. Enzymatic analysis of the endoglucanase activity using Carboxymethyl-Curdlan-Remazol Brilliant Blue (CM-Curdlan-RBB) as a substrate revealed a wide temperature optimum of 24-40°C, a pH optimum of 5.0-6.5 and a K(m) of 0.8 mg ml(-1). HPAEC analysis of the products formed by Eng2 when acting on different oligo-β-1,3-glucans confirmed the predicted endoglucanase activity and also revealed a transferase activity for oligosaccharides of a low degree of polymerization. The growth phenotype of the Afeng2 mutant was identical to that of the wt strain.
Hartl
Lukas
L
Unité des Aspergillus, Département de Parasitologie et Mycologie, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris cedex 15, France.
Gastebois
Amandine
A
Aimanianda
Vishukumar
V
Latgé
Jean-Paul
JP
D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D004795:Enzyme Stability; D017353:Gene Deletion; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature
D011994:Recombinant Proteins; D004693:Glucan Endo-1,3-beta-D-Glucosidase