Zhuravleva L I, Oreshkin E N, Bezborodov A M
Prikl Biokhim Mikrobiol. 1987 Mar-Apr;23(2):208-15.
A procedure for isolation and purification of restriction endonuclease Sac I from Streptomyces achromogenes ATCC 12767 is proposed. It allows to obtain an electrophoretically homogeneous enzyme preparation with the purification degree 1097 and the enzyme yield by activity 3.7%. The molecular weight of SacI was found to be 52,000 +/- 5,000 D, and isoelectric point 6.2. The enzyme consists of two subunits, which was found by polyacrylamide gel electrophoresis under denaturing conditions. Km and Vmax values were determined for the enzymatic reaction; they are equal to 4.6 X 10(-9) M and 9.19 X 10(-10) M/min, respectively.
本文提出了一种从无色链霉菌ATCC 12767中分离纯化限制性内切酶Sac I的方法。该方法可获得电泳纯的酶制剂，纯化度为1097，酶活性产率为3.7%。SacI的分子量为52,000±5,000 D，等电点为6.2。在变性条件下通过聚丙烯酰胺凝胶电泳发现该酶由两个亚基组成。测定了酶促反应的Km和Vmax值；它们分别等于4.6×10(-9) M和9.19×10(-10) M/min。
