Purification and Characterization of Mannose Isomerase from Agrobacterium radiobacter M-1
Purification and Characterization of Mannose Isomerase from Agrobacterium radiobacter M-1
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DOI：
10.1271/bbb.65.658
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发表时间：
2001-01
期刊：
影响因子：
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通讯作者：
J. Hirose;K. Maeda;H. Yokoi;Y. Takasaki
中科院分区：
文献类型：
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作者：
J. Hirose;K. Maeda;H. Yokoi;Y. Takasaki
从放射性土壤杆菌M-1（以前的假单胞菌属MI）的甘露糖异构酶纯化到电泳同质性和特征。无细胞提取物通过硫酸铵分级、Butyl-Toyopolymer 650 M、DEAE-Sepharose和羟基磷灰石柱层析分离。其分子量通过SDS-PAGE估计为44 kDa，通过凝胶过滤估计为90 kDa，其中该酶很可能是由两个相同亚基组成的二聚体。纯化的酶具有8.0的最适pH、60°C的最适温度、5.2的pI和20 mM的Km，并且特异性地将D-甘露糖和D-来苏糖转化为酮糖。鉴定了N-末端氨基酸序列。
A mannose isomerase from Agrobacterium radiobacter M-1 (formerly Pseudomonas sp. MI) was purified to electrophoretic homogeneity and characterized. A cell-free extract was separated by ammonium sulfate fractionation, Butyl-Toyopearl 650M, DEAE-Sepharose and hydroxylapatite column chromatography. Its molecular mass was estimated to be 44 kDa by SDS-PAGE and 90 kDa by gel filtration, in which the enzyme is most likely a dimer composed of two identical subunits. The purified enzyme had an optimum pH at 8.0, an optimum temperature at 60°C, a pI of 5.2 and a Km of 20 mM, and specifically converted D-mannose and D-lyxose to ketose. The N-terminal amino acid sequence was identified.