Extracellular nuclease produced by a marine bacterium. II. Purification and properties of extracellular nuclease from a marine Vibrio sp.
Extracellular nuclease produced by a marine Vibrio sp., strain No. 2, was purified by salting out with ammonium sulfate and by chromatography on a DEAE-cellulose column and twice on a Sephadex G-200 column. The nuclease was eluted as a single peak in which the deoxyribonuclease (DNase) activity and ribonuclease (RNase) activity appeared together. Polyacrylamide disc gel electrophoresis showed a single band of stained protein which had both DNase and RNase activity. The molecular weight of the enzyme was estimated to be 100 000 daltons. When using partially purified enzyme from the DEAE-cellulose column, the optimum pH for activity was 8.0, and the enzyme was activated strongly by 0.05 M Mg2+ ions and stabilized by 0.01 M Ca2+ ion. These concentrations of Mg2+ and Ca2+ ions are similar to those of the two cations in seawater. Indeed, the enzyme revealed high activity and strong stability when kept in seawater. The presence of particulate matter, such as cellulose powder, chitin powder. Hyflosupercel, Kaolin, and marine mud increased the stability of the enzyme. When the hydrostatic pressure was increased from 1 to 1000 atmospheres, the decrements of the enzyme activity were more pronounced at 30 and 40 degrees C than at 25 or 50 degrees C. The enzyme activity was restored after decompression to 1 atm at 30 degrees C.
D002118:Calcium / Q000494:pharmacology; D003851:Deoxyribonucleases / Q000302:isolation & purification / Q000378:metabolism; D004355:Drug Stability; D005092:Exonucleases / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006874:Hydrostatic Pressure; D008274:Magnesium / Q000494:pharmacology; D008722:Methods; D008970:Molecular Weight; D012623:Seawater; D013696:Temperature; D014733:Vibrio / Q000201:enzymology*; D014871:Water Microbiology
D003851:Deoxyribonucleases; D005092:Exonucleases; D008274:Magnesium; D002118:Calcium