Purification and characterization of beta-N-acetylhexosaminidases and beta-galactosidase from Streptococcus 6646 K.
Purification and characterization of beta-N-acetylhexosaminidases and beta-galactosidase from Streptococcus 6646 K.
复制标题
链球菌 6646 K 中 β-N-乙酰氨基己糖苷酶和 β-半乳糖苷酶的纯化和表征。
DOI：
10.1093/oxfordjournals.jbchem.a131263
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发表时间：
1976
影响因子：
2.7
通讯作者：
T. Osawa
中科院分区：
文献类型：
--
作者：
T. Kiyohara;T. Terao;K. Shioiri‐Nakano;T. Osawa
通过在对氨基苯基β-D-硫代半乳糖苷取代的琼脂糖凝胶和N-（对氨基苯基）草氨酸取代的琼脂糖凝胶上组合柱层析，从Streptococcus 6646 K组的培养物滤液中纯化三种β-N-乙酰氨基己糖苷酶[EC 3.2.1.23]和一种β-半乳糖苷酶[EC 3.2.1.52]。这些β-N-乙酰氨基己糖苷酶在pH 5.0和5.5之间表现出最佳活性，并且可以水解合成底物和糖肽底物。糖脂如GM 2、去唾液酸GM 2和globoside I对这些β-氨基己糖苷酶不敏感。β-半乳糖苷酶纯化11，000倍以上，其底物特异性与E.杆菌EDTA对该酶有抑制作用，Mn ~（2+）、Ca ~（2+）、Mg ~（2+）对该酶有激活作用。本文还讨论了亲和层析在糖苷酶纯化中应用的有关问题。
Three beta-N-acetylhexosaminidases [EC 3.2.1.52] and one beta-galactosidase [EC 3.2.1.23] were purified from the culture filtrate of streptococcus 6646 group K by a combination of column chromatographies on p-aminophenyl beta-D-thiogalactopyranoside-substituted Sepharose and N-(paminophenyl)oxamic acid-substituted Sepharose. These beta-N-acetylhexosaminidases showed optimal activities between pH 5.0 and 5.5 and could hydrolyze synthetic and glycopeptidic substrates. Glycolipids such as GM2, asialo-GM2, and globoside I were no susceptible to these beta-hexosaminidases. beta-Galactosidase, which was purified more than 11,000-fold, had a substrate specificity rather similar to that of beta-galactosidase from E. coli. This enzyme was inhibited by EDTA and activated by Mn2+, Ca2+, and Mg2+. Problems pertinent to the application of affinity chromatography to the purification of glycosidases are also discussed.