Purification and characterization of an extracellular, uracil specific ribonuclease from a Bizionia species isolated from the marine environment of the Sundarbans.
The first ribonuclease (RNase) from the Cytophaga-Flavobacterium-Bacteroides phylum, dominant in the marine environment, and also from the first Bizionia species isolated from the tropics was purified and characterized. Extracellular RNase production occurred when the culture medium contained 5-7% (w/v) NaCl. The 53.0 kDa enzyme was purified 29 folds with a recovery of 4% and specific activity of 630unit/mg protein. The pH and temperature optima are 6.5 and 35 degrees C, respectively and the enzyme retains more than half of its activity (relative to optimal assay conditions) after 1h pre-incubation separately with 5% (w/v) NaCl or from pH 5.0 to 8.5 or at 50 degrees C. Dithiothreitol and beta-mercaptoethanol do not inhibit whereas human placental RNase inhibitor protein halves the RNase activity. While Mg(2+), Ba(2+) and Ca(2+) enhanced the enzyme activity, Fe(2+), Cu(2+) and Hg(2+) inactivated it. This RNase degrades uracil containing nucleic acids only. Our isolate could be a novel renewable source of deoxyribonuclease (DNase)--free RNase enzyme.
Sana
Barindra
B
Environmental Science Programme and Department of Life Science and Biotechnology, Jadavpur University, Kolkata, India.
Ghosh
Debashish
D
Saha
Malay
M
Mukherjee
Joydeep
J
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D017077:Culture Media, Conditioned / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D041965:Flavobacteriaceae / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D019015:Geologic Sediments / Q000382:microbiology*; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000494:pharmacology; D008386:Marine Biology; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D009792:Oceans and Seas; D011131:Polyribonucleotides / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D012260:Ribonucleases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D014498:Uracil; D014871:Water Microbiology
D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D004791:Enzyme Inhibitors; D007477:Ions; D008670:Metals; D011131:Polyribonucleotides; D014498:Uracil; D012313:RNA; D012260:Ribonucleases