Purification and characterization of cycloinulooligosaccharide fructanotransferase from Bacillus macerans CFC1
Purification and characterization of cycloinulooligosaccharide fructanotransferase from Bacillus macerans CFC1
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浸软芽孢杆菌 CFC1 环菊寡糖果糖转移酶的纯化和表征
DOI：
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发表时间：
1998
影响因子：
2.8
通讯作者：
Yong
中科院分区：
文献类型：
--
作者：
Hwa;Yong
从浸软芽孢杆菌CFC 1的培养液中将从菊粉产生环果聚糖的环菊寡糖果聚糖转移酶（CFTase）纯化332倍。通过SDS-聚丙烯酰胺凝胶电泳和凝胶过滤，估计CFT酶的分子量为110 kDa，表明该酶具有单体结构。在pH 7.5和45°C下观察到最大水平的酶活性。该酶在pH 6.0至9.5范围内稳定，并在高达45°C的温度下稳定1小时。当Ag +或Cu 2+浓度为0.5mM时，酶活性完全被抑制.蔗糖（GF），1-蔗果三糖（GF 2），或nystose（GF 3）被发现是底物的CFT酶，但大于nystose的菊粉寡糖被攻击的酶。CFTase不仅催化作为主要反应的环化，而且以与环糊精葡聚糖转移酶（CGTase）相同的方式催化涉及分子间转果糖基化的缩合和偶联反应（EC 2.4.1.19）。
Cycloinulooligosaccharide fructanotransferase (CFTase) which produces cyclofructan from inulin was purified 332-fold from a culture broth of Bacillus macerans CFC1. The molecular mass of the CFTase was estimated to be 110 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indicating that the enzyme has a monomer structure. The maximal level of enzyme activity was observed at pH 7.5 and 45°C. The enzyme was stable in the pH range 6.0 to 9.5, and at temperatures up to 45°C for 1 h. The enzyme activity was completely inhibited in the presence of 0.5 mM Ag + or Cu 2+ ion. None of sucrose (GF), 1-kestose (GF2), or nystose (GF3) were found to be substrates for the CFTase, but inulooligosaccharides larger than nystose were attacked by the enzyme. The CFTase catalyzes not only the cyclization as the major reaction, but also disproportionation and coupling reactions involving intermolecular transfructosylation in the same manner as cyclodextrin glucanotransferase (CGTase) (EC 2.4.1.19).