Purification and Properties of beta-N-Acetylhexosaminidase from Mucor fragilis Grown in Bovine Blood.
Mucor fragilis grown on bovine blood powder as the sole carbon source abundantly produced beta-N-acetylhexosaminidase. The enzyme activity was several times higher than that of a culture obtained with glucose medium. The enzyme had two different molecular weight forms. The high-molecular-weight form had somewhat higher beta-N-acetylgalactosaminidase activity than the lower-molecular-weight enzyme which had beta-N-acetylgalactosaminidase activity equivalent to about 40% of its beta-N-acetylglucosaminidase activity. Bovine blood seemed to induce both enzymes, but N-acetylamino sugars specifically induced the low-molecular-weight form. N-Acetylgalactosamine had an especially marked effect on activity. The low-molecular-weight form of enzyme was purified from the culture filtrate by fractionation with ammonium sulfate and various column chromatographies. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis. The optimum pH was 4.0 to 5.0 for beta-N-acetylglucosaminidase activity and 5.5 to 6.5 for beta-N-acetylgalactosaminidase activity. The enzyme hydrolyzed natural substrates such as di-N-acetylchitobiose, tri-N-acetylchitotriose, and a glycopeptide obtained by modification of fetuin.
Yamamoto
K
K
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Kyoto 606, and Institute of Soy Sauce, Higashimaru Shoyu Co. Ltd., Tatsuno, Hyogo 679-41, Japan.
Tsuji
Y
Y
Matsushita
S
S
Kumagai
H
H
Tochikura
T
T