A restriction endonuclease (DpaI) from Deleya pacifica IAM 14115, a marine bacterium, an isoschizomer of ScaI.

physicochemical properties. Deleya pacifica (Baumann et al. 1972) Baumann et al. 1983 IAM 141 15*1 was aerobically grown for 24hr at 30°C in a modified marine broth as described previously.2) The cells (30g) harvested from 6liters of culture broth by centrifugation were suspended in 200ml of a sonication buffer (pH 7.5) containing 10mM K2HPO4, 7mM 2-mercaptoethanol, 100mM NaCl and 20/xg/ml phenylmethylsulfonyl fluoride (w/v), and disrupted by sonication (9kHz) at 200W for 6min. The sonicated cell suspension was centrifuged for 60min at 27,000 x g. The resulting supernatant (210ml) was put onto a phosphocellulose column (Whatman P-l l, 3.0 x 14.0cm, 100 ml). The column was developed with a linear gradient of buffer A (10mM K2HPO4, 2-mercaptoethanol, 0.1 m NaCl and 5% glycerol, v/v, pH 7.5) containing 0.1 to 1.0m NaCl (1,000ml in total) at a flow rate of 50ml/hr. The enzyme was recovered in fractions 14-30 (10ml each,