Purification and characterization of beta-N-acetylhexosaminidase from Trichoderma harzianum.
beta-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 degrees C, respectively. The enzyme hydrolyzed N-acetylchitooligosaccharides at the non-reducing ends to release GlcNAc monomer. The enzyme showed a strict substrate specificity to the sugar chains in complex carbohydrates, hydrolyzing only the linkage of GlcNAc beta 1-3Gal, but not hydrolyzing the other linkages such as GalNAc beta 1-3Gal and GlcNAc beta 1-2Man.
Koga
K
K
Biological Engineering Laboratory, Asahi Industries Co., Ltd., Saitama, Japan.
Iwamoto
Y
Y
Sakamoto
H
H
Hatano
K
K
Sano
M
M
Kato
I
I
D000116:Acetylgalactosamine / Q000031:analogs & derivatives / Q000378:metabolism; D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000378:metabolism; D002240:Carbohydrate Sequence; D004795:Enzyme Stability; D005456:Fluorescent Dyes; D005656:Fungal Proteins / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology*; D001619:beta-N-Acetylhexosaminidases / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism
D005456:Fluorescent Dyes; D005656:Fungal Proteins; D009844:Oligosaccharides; C014123:4-nitrophenyl-2-acetamido-2-deoxygalactopyranoside; C014124:4-nitrophenyl-N-acetyl-2-deoxyglucopyranoside; D001619:beta-N-Acetylhexosaminidases; D000116:Acetylgalactosamine; D000117:Acetylglucosamine