Purification and Characterization of 4N-Trimethylamino-1-butanol Dehydrogenase of Pseudomonas sp . 13 CM
Purification and Characterization of 4N-Trimethylamino-1-butanol Dehydrogenase of Pseudomonas sp . 13 CM
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假单胞菌 4N-三甲氨基-1-丁醇脱氢酶的纯化和表征。
DOI：
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发表时间：
2007
期刊：
影响因子：
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通讯作者：
N. Mori
中科院分区：
文献类型：
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作者：
Maizom Hassan;S. Morimoto;H. Murakami;Tsuyoshi Ichiyanagi;N. Mori
假单胞菌的一种新酶--依赖NAD的4-N-三甲氨基-1-丁醇脱氢酶13 cm，经5步层析纯化526倍，均一。该酶的相对分子质量为45 kDa，为单体酶。测得等电点为4.8。最佳反应温度为50℃，氧化反应和还原反应的最佳pH值分别为9.5和6.0。对纯化的酶进行了底物专一性、动力学参数和氨基酸末端序列的进一步表征。三甲氨基-1-丁醇和NAD-2的Km值分别为0.54 mM和0.22 mM。在还原反应中，三甲氨基丁醛和NADH的表观Km值分别为0.67 mM和0.04 mM。该酶受SH试剂、络合剂和重金属离子的抑制。对N末端12个氨基酸残基进行了测序。
A new enzyme, NADþ-dependent 4-N-trimethylamino-1-butanol dehydrogenase from Pseudomonas sp. 13CM, was purified 526-fold to apparent homogeneity in 5 chromatographic steps. The enzyme had a molecular mass of 45 kDa and appeared to be a monomer enzyme. The isoeletric point was found to be 4.8. The optimum temperature was 50 C, and the optimum pHs for the oxidation and reduction reactions were 9.5 and 6.0 respectively. The purified enzyme was further characterized with respect to substrate specificity, kinetic parameters, and amino acid terminal sequence. The Km values for trimethylamino-1-butanol and NADþ were 0.54mM and 0.22mM respectively. In the reduction reaction, the apparent Km values for trimethylaminobutylaldehyde and NADH were 0.67mM and 0.04mM, respectively. The enzyme was inhibited by SH reagents, chelating reagents, and heavy metal ions. The Nterminal 12 amino acid residues were sequenced.