An Intracellular Endonuclease of Bacillus subtilis Specific for Single‐Stranded DNA

• Published in: European journal of biochemistry Vol. 61; no. 2; pp. 487 - 492
• Main Authors: CIARROCCHI, Giovanni, FORTUNATO, Adriana, COBIANCHI, Fabio, FALASCHI, Arturo
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.01.1976
• Subjects: Bacillus subtilis - enzymology; Calcium - pharmacology; Chloromercuribenzoates - pharmacology; Deoxyribonucleases - isolation & purification; Deoxyribonucleases - metabolism; Endonucleases - isolation & purification; Endonucleases - metabolism; Enzyme Activation - drug effects; Hydrogen-Ion Concentration; Isoenzymes - analysis; Isoenzymes - metabolism; Kinetics; Magnesium - pharmacology; Molecular Weight; Osmolar Concentration; Potassium Chloride - pharmacology
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1111/j.1432-1033.1976.tb10043.x

We have fractionated from extracts of Bacillus subtilis the DNase activity specific for singlestranded DNA; the activity separates in two main fractions on Sephadex G‐200, a larger one (Mr < 400000) and a smaller one (Mr∼ 30000). We have purified the smaller, more abundant fraction nearly 3000‐fold. The purified enzyme has a pH optimum close to 8, is activated by Ca2+, and is inhibited by EDTA; the enzyme hydrolyses single‐stranded DNA at a rate approximately 40 times greater than double‐stranded DNA. The mode of action is endonucleolytic on both substrates. but the possibility that the two activities may reside on different molecules is not ruled out. The products have 5′‐P and 3′‐OH ends. The enzyme is different from those purified from the culture media of the same organism in several respects; the latter are all extracellular enzymes, they are not specific for single‐stranded DNA (except one) and have all an exonucleolytic mode of action.