4-N-三甲基氨基丁醛脱氢酶：来自假单胞菌属13CM的一种酶的纯化及特性研究
4-N-trimethylaminobutyraldehyde dehydrogenase: purification and characterization of an enzyme from Pseudomonas sp. 13CM.
作者信息
Hassan Maizom, Okada Masahiro, Ichiyanagi Tsuyoshi, Mori Nobuhiro
机构信息
United Graduate School of Agricultural Sciences, Tottori University, Koyama, Tottori, Japan.
出版信息
Biosci Biotechnol Biochem. 2008 Jan;72(1):155-62. doi: 10.1271/bbb.70514. Epub 2008 Jan 7.
4-N-trimethylaminobutyraldehyde dehydrogenase from Pseudomonas sp. 13CM was purified 14-fold to apparent homogeneity by hydrophobic chromatography on a Phenyl-Toyopearl, and affinity chromatography was done on a 5'-AMP Sepharose4B in the presence of dithiothreitol. The enzyme was found to be a trimer with identical 55 kDa subunits. The isoeletric point was found to be 5.5. The optimum temperature and pH were 40 degrees C and pH 10.0. The purified enzyme was further characterized with respect to substrate specificity, kinetic parameters, and analog inhibition. The K(m) values for 4-N-trimethylaminobutyraldehyde, 4-dimethylaminobutyraldehyde, and NAD(+) were 7.4, 51, and 125 microM respectively. The enzyme was inhibited by SH reagents, and by heavy metal ions.
从假单胞菌属13CM中纯化得到的4-N-三甲基氨基丁醛脱氢酶，通过在苯基-琼脂糖凝胶上进行疏水层析纯化了14倍，达到表观均一性，并且在二硫苏糖醇存在的条件下，在5'-AMP琼脂糖凝胶4B上进行亲和层析。该酶被发现是由相同的55 kDa亚基组成的三聚体。其等电点为5.5。最适温度和pH分别为40℃和pH 10.0。对纯化后的酶在底物特异性、动力学参数和类似物抑制方面进行了进一步表征。4-N-三甲基氨基丁醛、4-二甲基氨基丁醛和NAD(+)的K(m)值分别为7.4、51和125 microM。该酶受到巯基试剂和重金属离子的抑制。
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