Streptomyces sp.No.499によって生産されるβ-N-アセチルヘキソサミニダーゼの精製と性質

概要

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In the course of our screening program for chitinase and β-N-acetylhexosaminidase (HexoNAcase) from the fermented broth of about 800 stock cultures of Streptomyces sp. isolated from soil samples, the highest amounts of both enzymes were found to be produced in the broth by Streptomyces sp. No. 499 isolated from a soil sample collected at Toyonaka-shi, Osaka, Japan. The enzyme was produced by Streptomyces sp. No. 499 cultivated with chitin as the growth substrate. HexoNAcase from the filtrate of the broth was purified by chromatography using, successively, Amberlite CG-50,P-cellulose, and Sephadex G-100 columns. The enzyme was completely purified, its purity being comfirmed by SDS-polyacrylamide gel electrophoresis (PAGE). The molecular weight of the enzyme was demonstrated to be 52,000 by a gel chromatography and 57,000 by SDS-PAGE. Its optimum pH and temperature for hydrolytic action were 6.0 and 55℃ towards p-Nitrophenyl-β-N-acetylglucosaminide, and 5.0 and 55℃ towards p-Nitrophenyl-β-N-acetylgalactosaminide. The enzyme was confirmed to readily hydrolyze chitobiose, chitotriose, and chitotetriose in that order, releasing N-acetylglucosamine in the mode of the exo-type hydrolase.
社団法人日本生物工学会の論文
1998-05-25