Partial purification of a thermostable dextranase using Sephacryl S‐300 adsorption
Partial purification of a thermostable dextranase using Sephacryl S‐300 adsorption
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使用 Sephacryl S-300 吸附部分纯化热稳定性葡聚糖酶
DOI：
10.1046/j.1472-765x.1997.00238.x
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
C. Wynter
中科院分区：
文献类型：
--
作者：
C. Wynter
来自新西兰温泉中分离的嗜热厌氧菌株 Rt364 的耐热葡聚糖酶（1,6-α-d-葡聚糖 6-葡聚糖水解酶）通过吸附到基于葡聚糖的色谱树脂 Sephacryl S-300 上，从无细胞上清液中部分纯化。用 2% T10 葡聚糖竞争性洗脱、透析、浓缩并通过 SDS-PAGE 检查。总体回收率为 47%，此过程的比活性增加了 25 倍。此前发现 Rt364 葡聚糖酶的最佳温度为 80 °C，可水解 α-1,6 和 α-1,4 糖苷键。 Sephacryl S-300 吸附是一个简单、有用的步骤，普遍应用于浓缩和纯化水解葡聚糖的细菌酶。
Thermostable dextranase (1,6‐α‐d‐glucan 6‐glucanohydrolysase) from a thermophilic anaerobic bacterium strain Rt364, isolated from a New Zealand hot spring, was partially purified from the cell‐free supernatant fluid by adsorption onto Sephacryl S‐300, a dextran‐based chromatographic resin. It was competitively eluted with 2% T10 dextran, dialysed, concentrated and examined by SDS–PAGE. The overall recovery was 47% and the increase in specific activity by this procedure was 25‐fold. The Rt364 dextranase had previously been found to have an optimum temperature of 80 °C and hydrolysed both α‐1,6 and α‐1,4 glucosidic bonds. Sephacryl S‐300 adsorption is a simple, useful step with general application for concentrating and purifying bacterial enzymes that hydrolyse dextrans.