Streptolysin S: improved purification and characterization.
Streptolysin S: improved purification and characterization.
复制标题
链球菌溶血素 S：改进的纯化和表征。
DOI：
10.1016/0003-9861(78)90423-x
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发表时间：
1978
影响因子：
3.9
通讯作者：
T. Akao
中科院分区：
文献类型：
--
作者：
Chun;M.;J. Faría;T. Akao
研究了一种简单高效的产纯化链溶素S的方法。溶血素的最大产量发生在快速生长的链球菌到达固定期1小时后收获，并与诱导剂低核糖核苷酸孵育。在0.1min 50 mm磷酸钾缓冲液（pH 6.8）条件下，发现乙酸铵能有效地保护链溶素S免于热失活，并在整个纯化过程中使用缓冲液。经三步纯化后的产物具有最高的比活性（1.2 × 107HU / mg蛋白），回收率为35% ~ 45%。估计链溶素S和载体寡核苷酸的表观分子量分别为15,000和7,100。通过研究各种水解酶对溶血链蛋白酶S的作用，证实了其活性原理的多肽性质；只有蛋白酶、凝乳胰蛋白酶和枯草菌素能灭活溶血素。氨基酸分析表明，活性肽由32个氨基酸残基组成。
A simple and efficient procedure for the production and purification of streptolysin S has been developed. Maximal production of the hemolysin occurred when rapidly grown streptococci were harvested 1 h after reaching the stationary phase and incubated with the inducer oligoribonucleotide. Ammonium acetate at 0.1min 50 mmpotassium phosphate buffer, pH 6.8, was found to effectively protect streptolysin S from thermal inactivation, and was used in the buffer throughout purification. The three-step purification procedure resulted in preparations with the highest specific activity (1.2 × 107HU per mg protein) ever reported, in recoveries ranging from 35 to 45%. The apparent molecular weights of streptolysin S and the carrier oligonucleotide were estimated as 15,000 and 7,100, respectively. The peptide nature of the active principle was confirmed by studies of the effects of various hydrolytic enzymes on streptolysin S; only pronase, chymotrypsin and subtilisin were found to inactivate the hemolysin. Amino acid analyses indicated that the active peptide consisted of 32 amino acid residues.