Isolation of deoxyribonuclease II from bovine intestinal mucosa

Mucosa from bovine small intestine was homogenized in Krebs–Ringer phosphate buffer, pH 7.8, the homogenate centrifuged at 16 300 × g, and the supernatant solution filtered through cheesecloth to remove lipid material. The filtrate was centrifuged at 105 000 × g and the supernatant solution chromatographed on DEAE-cellulose. The major peak of DNase II activity, eluted with 20 mM phosphate – 10 mM EDTA buffer, pH 7.8, was purified further by ion-exchange chromatography on CM-cellulose and gel filtration on Sephadex G-100. The enzyme was purified 78-fold in 13% yield. Evidence was adduced to indicate that the second minor peak of DNase II activity, eluted from the DEAE-cellulose by a potassium chloride gradient in the 20 mM phosphate – 10 mM EDTA buffer, was an artifact arising from the presence of significant amounts of DNA in the 105 000 × g supernatant. The enzyme degraded DNA endonucleolytically to 3′-PO4, 5′-OH oligonucleotides and is similar in its properties to DNase II from other tissues.