Arutiunian E E, Gruber I M, Poliachenko V M, Nikol'skaia I I, Debov S S
Vopr Med Khim. 1990 Mar-Apr;36(2):75-9.
Heterogeneous profile of methylases was found in S. aureus 6782 strain. A procedure was developed for isolation of individual methylases from Sau 6782 free of Sau 6782 restrictases and unspecific nucleases by means of hydrophobic chromatography on phenyl-Sepharose. Ion exchange, affinity chromatographies and gel filtration were also used for isolation of the Sau 6782 methylases. But this technique was of limited suitability for isolation of individual methylating enzymes of the Sau 6782 type because it did not allow to separate completely these methylases from contaminating enzymes of DNA degradation. Effects of salts, glycerol and Triton X-100 on activity of total preparation of methylases Sau 6782 were studied. Na+ and K+ chlorides, ammonium sulfate at concentrations 0.4 M and highen as well as Triton X-100 inhibited reversibly the methylase activity followed by complete reduction up to initial level after dialysis. Glycerol at concentration 60% activated Sau 6782 methylases by 50% and stabilized the enzyme.
在金黄色葡萄球菌6782菌株中发现了甲基化酶的异质性谱。开发了一种从Sau 6782中分离单个甲基化酶的方法，通过在苯基琼脂糖上进行疏水色谱法，可去除Sau 6782限制性内切酶和非特异性核酸酶。离子交换、亲和色谱和凝胶过滤也用于分离Sau 6782甲基化酶。但该技术对分离Sau 6782型单个甲基化酶的适用性有限，因为它无法将这些甲基化酶与DNA降解的污染酶完全分离。研究了盐、甘油和Triton X-100对Sau 6782甲基化酶总制剂活性的影响。0.4 M及更高浓度的氯化钠、氯化钾、硫酸铵以及Triton X-100可逆性抑制甲基化酶活性，透析后活性完全恢复至初始水平。60%浓度的甘油可使Sau 6782甲基化酶活性提高50%并使酶稳定。
