Kovalevskaia N P, Zelinskaia N V, Zheleznaia L A, Matvienko N I
Bioorg Khim. 1993 Nov;19(11):1073-6.
New site-specific endonucleases BspBS31I, BstBS32I, BspIS41, BstTS5I, BspTS514I were isolated from five thermophilic soil bacteria Bacillus sp. BS31, B. stearothermophilus BS32, Bacillus sp. IS4, B. stearothermophilus TS5, Bacillus sp. TS514. The enzymes are isoschizomers of the restriction endonuclease BbvII. Endonuclease BspTS514I was obtained pure from interfering contaminations by two consecutive chromatographies on blue agarose and hydroxyapatite. The enzyme exhibits a maximal activity at 55 degrees C in 10 mM tris-HCl (pH 9.2), 10 mM MgCl2 and 50 mM NaCl.
新型位点特异性内切酶BspBS31I、BstBS32I、BspIS41、BstTS5I、BspTS514I是从五株嗜热土壤细菌芽孢杆菌属BS31、嗜热脂肪芽孢杆菌BS32、芽孢杆菌属IS4、嗜热脂肪芽孢杆菌TS5、芽孢杆菌属TS514中分离得到的。这些酶是限制性内切酶BbvII的同裂酶。通过在蓝色琼脂糖和羟基磷灰石上连续两次色谱法,从干扰性污染物中获得了纯的内切酶BspTS514I。该酶在10 mM三羟甲基氨基甲烷盐酸盐(pH 9.2)、10 mM氯化镁和50 mM氯化钠中,于55℃时表现出最大活性。