Purification and characterization of an extracellular beta-n-acetylhexosaminidase from Paecilomyces persicinus

Abstract

Both beta-N-acetylglucosaminidase nad beta-N-acetylgalactosaminidase activities were detected in the culture fluids of Paecilomyces persicinus P-10 after growth in a soybean meal-corn meal medium. The active material was purified by means of protamine sulfate fractionation and ultrafiltration, followed by ion exchange and gel chromatography. The ratio of the two activities remained constant throughout the purification, and the final product was shown to migrate as a single band by using gel isoelectric focusing, disc electrophoresis, and detergent gel electrophoresis. Temperature, pH, inhibition, and kinetic studies were performed to characterize both activities. The molecular weight of the enzyme was estimated to be about 100,000 by high-resolution gel chromatography. Based on the data obtained, it is suggested that both beta-N-acetylglucosaminidase and beta-N-acetylgalactosaminidase activities reside in the same protein.

Purification and characterization of ββ‐N‐acetylhexosaminidase from the phytopathogenic fungusBipolaris sorokiniana

“…The enzyme b-N-acetylhexosaminidase was purified from the fungus B. sorokiniana. Purification was about 70-fold, yielding 41%, similar to the data described for other microorganisms (Eriquez and Pisano 1979;Jones and Kosman 1980;Nanjo et al 1990). The purification step of gel filtration was effective where the major quantity of protein eluted after the enzyme.…”

“…The enzyme β‐N‐acetylhexosaminidase was purified from the fungus B. sorokiniana . Purification was about 70‐fold, yielding 41%, similar to the data described for other micro‐organisms (Eriquez & Pisano 1979; Jones & Kosman 1980; Nanjo et al . 1990).…”

Purification and Properties of β- N -Acetylhexosaminidase from Mucor fragilis Grown in Bovine Blood

“…The properties of the enzyme from M. fragilis are not significantly different from those reported for this enzyme from other fungi. However, the enzyme was not inhibited by GIcNAc or GalNAc alone or together; these results are different from those for the ,B-N-acetylhexosaminidases of other fungi reported previously (5,11,12,17). The enzyme Effects of pH on activity and stability of P-N-acetylhexosaminidase.…”

“…little or no effect on enzyme activity. Neither ,-GlcNAcase nor P-GalNAcase activity was inhibited by GIcNAc or GalNAc, unlike other mold enzymes (5,11,12,17).…”

Comparison of chitinolytic enzymes from an alkaline, hypersaline lake and an estuary

“…1C). GlcNAcidase activity has been observed over a pH range of 2-10 in Paecilomyces persicinus; however, the level of activity at pH 10 was approximately 5% of its optimal activity at pH 5.8 (Eriquez and Pisano, 1979). Keyhani and Roseman (1996) found a periplasmic GlcNAcidase from Vibrio furnissii to have a pH optimum approaching 7 with substrates (GlcNAc) 3-6.…”