从嗜盐杆菌 Alk36 中克隆、纯化和表征重组嘌呤核苷磷酸化酶。
Cloning, purification and characterisation of a recombinant purine nucleoside phosphorylase from Bacillus halodurans Alk36.
机构信息
Enzyme Technologies Group, CSIR Biosciences, Private Bag X2, Modderfontein, Johannesburg, 1645, South Africa.
出版信息
Extremophiles. 2010 Mar;14(2):185-92. doi: 10.1007/s00792-009-0297-4.
Abstract
A purine nucleoside phosphorylase from the alkaliphile Bacillus halodurans Alk36 was cloned and overexpressed in Escherichia coli. The enzyme was purified fivefold by membrane filtration and ion exchange. The purified enzyme had a V (max) of 2.03 x 10(-9) s (-1) and a K (m) of 206 microM on guanosine. The optimal pH range was between 5.7 and 8.4 with a maximum at pH 7.0. The optimal temperature for activity was 70 degrees C and the enzyme had a half life at 60 degrees C of 20.8 h.
摘要
从嗜碱芽孢杆菌 Alk36 中克隆并在大肠杆菌中过表达了一种嘌呤核苷磷酸化酶。该酶通过膜过滤和离子交换进行了五倍纯化。纯化后的酶在鸟苷上的 V(max)为 2.03 x 10(-9) s(-1)，K(m)为 206 microM。最佳 pH 范围在 5.7 到 8.4 之间，在 pH 7.0 时达到最大值。活性的最佳温度为 70 摄氏度，该酶在 60 摄氏度下的半衰期为 20.8 小时。
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