A novel restriction endonuclease, Bsp 1894I, has been isolated from Bacillus sphaericus 1894, and its catalytic properties have been studied. For the purification of this enzyme, 35g of cells were used and disrupted by French Press at 20, 000 P.S.I. After streptomycin sulfate and ammonium sulfate fractionation, DEAE-cellulose, QAE-Sephadex, hydroxyapatite, and Sephadex G-100 chromatography have been carried out. From the dideoxy sequencing and the comparison of digestion pattern on various DNAs, it was demonstrated that the new enzyme Bsp 1894I recognizes 5'-G↓GNCC-3' and cleaves at the site indicated by the arrow. This enzyme is an isoschizomer of Asu I, Cfr 13I, Nsp 7524IV, and Sau 96I. In catalytic properties, Bsp 1894I shows maximum activity at range between 6 and 7 in the presence of 10 mM MgCl2. The optimum temperature of Bsp 1894I is 42℃. Unlike above isoschizomer, Bsp 1894I does not require NaCl for activity.
Bacillus sphaerious 1894 균주에서 새로운 제한효소 Bsp 1894I을 정제하였다. 35 g 의 세포를 French Press 로 깨고 streptomycin sulfate 와 ammonium sulfate 로 분획침전을 시킨다음, DEAE-cellulose, QAE-Sephadex, Hydroxyapatite, Gel permeation(G-100) 등 크로마토그래피를 거쳐 0.86 mg 의 enzyme 을 얻었다. dideoxy sequencing 과 다양한 DNA 에 대한 절단모형으로부터 이 Bsp 1894I 은 5'-G↓GNCC-3' 을 인지하고 화살표 부위를 자르는 효소임을 밝혀냈다. 효소적 특성에서 Bsp 1894I 은 10 mM MgCl2 와 pH6-7 에서 최대 활성을 보였다. 그리고 최적온도는 42℃ 였으며 활성을 위해 NaCl 을 요구하지 않았다.
