title: Halophilic Nuclease of a Moderately Halophilic Bacillus sp.: Production, Purification, and Characterization.
abstract: A moderately halophilic bacterium, Bacillus sp., isolated from rotting wood on the seashore in Nauru, produced an extracellular nuclease when cultivated aerobically in media containing 1 to 2 M NaCl. The enzyme was purified from the culture filtrate to an electrophoretically homogeneous state by ethanol precipitation, DEAE-Sephadex A-50 column chromatography, and Sephadex G-200 gel filtration. The enzyme consisted of two charge isomers and showed both RNase and DNase activities. Molecular weight was estimated to be 138,000 by Sephadex G-200 gel filtration. The enzyme had marked halophilic properties, showing maximal activities in the presence of 1.4 to 3.2 M NaCl or 2.3 to 3.2 M KCl. The enzyme hydrolyzed thymidine-5'-monophosphate-p-nitrophenyl ester at a rate that increased with NaCl concentration up to 4.8 M. In the presence of both Mg and Ca, activity was greatly enhanced. The activity was lost by dialysis against water and low-salt buffer, but it was protected when 10 mM Ca was added to the dialysis buffer. When the inactivated enzyme was dialyzed against 3.5 M NaCl buffer as much as 68% of the initial activity could be restored. The enzyme exhibited maximal activity at pH 8.5 and at 50 degrees C on DNA and at 60 degrees C on RNA and attacked RNA and DNA exonucleolytically and successively, producing 5'-mononucleotides.
authors.affiliation: Department of Agricultural Chemistry, Kagoshima University, Korimoto, Kagoshima-shi, Japan 890.
authors.forename: H
authors.initials: H
authors.lastname: Onishi
authors.forename: T
authors.initials: T
authors.lastname: Mori
authors.forename: S
authors.initials: S
authors.lastname: Takeuchi
authors.forename: K
authors.initials: K
authors.lastname: Tani
authors.forename: T
authors.initials: T
authors.lastname: Kobayashi
authors.forename: M
authors.initials: M
authors.lastname: Kamekura