Miller R L, Varner H H
Biochemistry. 1979 Dec 25;18(26):5828-32. doi: 10.1021/bi00593a026.
Lysyl hydroxylase from fetal porcine skin is shown to bind in a highly specific manner to aminoethyl-Sepharose 4B. When coupled to ammonium sulfate fractionation and DEAE-cellulose chromatography, chromatography of lysyl hydroxylase preparations on aminoethyl-Sepharose 4B has yielded a highly purified (greater than 95%) preparation of lysyl hydroxylase. The enzyme consists of two subunits with molecular weights of 70 000 and 115 000. The overall recovery of activity was 2.5%, yielding approximately to 3.5 mg of purified enzyme from 900 g of fetal porcine skin. The enzyme is more active at 30 degrees C than at 37 degrees C and has a pH optimum near 8.0. Both catalase and bovine serum albumin are required by the enzyme for maximum activity. The sulfhydryl reagents p-(chloromercuri)-benzoate, N-ethylmaleimide, and iodoacetamide are potent inhibitors of the enzyme, whereas dithiothreitol appears to be an activator.
已证明来自胎猪皮肤的赖氨酰羟化酶能以高度特异性的方式与氨乙基-琼脂糖4B结合。当与硫酸铵分级分离和DEAE-纤维素色谱法相结合时，在氨乙基-琼脂糖4B上对赖氨酰羟化酶制剂进行色谱分析可得到高度纯化（大于95%）的赖氨酰羟化酶制剂。该酶由两个亚基组成，分子量分别为70000和115000。活性的总回收率为2.5%，从900克胎猪皮肤中可得到约3.5毫克的纯化酶。该酶在30℃时比在37℃时更具活性，最适pH接近8.0。过氧化氢酶和牛血清白蛋白都是该酶发挥最大活性所必需的。巯基试剂对氯汞苯甲酸、N-乙基马来酰亚胺和碘乙酰胺是该酶的有效抑制剂，而二硫苏糖醇似乎是一种激活剂。
