[改进的从鸡心脏细胞溶质中纯化天冬氨酸转氨酶的方法。该酶的特性]
[Improved procedure for purification of aspartate transaminase from chicken heart cytosol. Characterization of the enzyme].
作者信息
Kochkina V M, Azarian A V, Mekhanik M L, Zakomyrdina L N, Sinitsyna N I
出版信息
Biokhimiia. 1978 Aug;43(8):1478-84.
The purification procedure reported includes fractionation of water extract from chicken hearts with ammonium sulfate, fractional precipitation with ethanol, chromatography on Whatman CM-52 cellulose and crystallization. Specific activity of the pure crystalline enzyme was 234 micromoles.min-1.mg-1, as determined in the coupled assay with malate dehydrogenase (pH 7.5; 25 degrees). The amino acid composition of the enzyme was determined and the circular dichroism spectrum was recorded in the 200-250 nm range. The spectrum shows two negative bands with extrema at 208 and 220 nm. From the circular dichroism data it is estimated that aspartate transaminase contains approximately 40% alpha-helix and 10% beta-structure.
所报道的纯化步骤包括用硫酸铵对鸡心水提取物进行分级分离、用乙醇分级沉淀、在Whatman CM - 52纤维素上进行色谱分离以及结晶。在与苹果酸脱氢酶的偶联测定中（pH 7.5；25摄氏度）测定，纯结晶酶的比活性为234微摩尔·分钟⁻¹·毫克⁻¹。测定了该酶的氨基酸组成，并记录了200 - 250纳米范围内的圆二色光谱。该光谱在208和220纳米处有两个极值的负带。根据圆二色性数据估计，天冬氨酸转氨酶含有约40%的α - 螺旋和10%的β - 结构。
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