Purification and Characterization of β-N-Acetylhexosaminidase from Trichoderma harzianum

Abstract

β-N-Acetylhexosaminidase was produced by Trichoderma harzianum cultivated with chitin as the growth substrate. The enzyme was purified 13.2-fold to homogeneity by ultrafiltration and sequential chromatography on SP-Toyopearl and Sephacryl S-200. The molecular weight of the enzyme was estimated to be about 150,000 by gel filtration. The pH and temperature optima were 4.0—5.5 and 50°C, respectively. The enzyme hydrolyzed N-acetylchitooligosaccharides at the nonreducing ends to release GlcNAc monomer. The enzyme showed a strict substrate specificity to the sugar chains in complex carbohydrates, hydrolyzing only the linkage of GlcNAcβ1-3Gal, but not hydrolyzing the other linkages such as GalNAcβ1-3Gal and GlcNAcβ1-2Man. © 1991, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.