Purification, Properties, and Transglycosylation Reaction of β-N-Acetylhexosaininidase from Nocardia orientalis(Biological Chemistry)

概要

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β-N-Acetylhexosaminidase (EC 3.2.1.52) from the culture filtrate of Nocardia orientalis was purified to homogeneity by precipitation with ammonium sulfate followed by column chromatography on CM-Sephadex, Bio-Gel P-60, and phenyl-Sepharose CL-4B. The molecular weight of the enzyme was about 56,000 by gel filtration and 54,000 by SDS polyacrylamide gel electrophoresis. The enzyme showed about 1.6-fold higher β-N-acetylglucosaminidase activity than β-N-acetylgalactosaminidase activity. The optimum pH and temperature were 5.0 and around 70-75℃ for PNP-GlcNAc, and 4.0 and 60℃ for PNP-GalNAc. The enzyme was stable in the pH range from 4.0 to 8.0 and below 45℃. The enzyme hydrolyzed N-acetyl-chitooligosaccharides, di-N-acetyl-chitobiose through hexa-N-acetyl-chitohexaose. The enzyme showed glycosyl transferase activity during the hydrolysis of di-N-acetyl-chitobiose. Two major transfer products were isolated and identified as the β-(1→6)-linked disaccharide of N-acetylglucosamine and tri-N-acetyl-chitotriose.
社団法人日本農芸化学会の論文
1990-04-23