title: Purification and characterization of a cold-adapted uracil-DNA glycosylase from Atlantic cod (Gadus morhua).
abstract: Uracil-DNA glycosylase (UDG; UNG) has been purified 17000-fold from Atlantic cod liver (Gadus morhua). The enzyme has an apparent molecular mass of 25 kDa, as determined by gel filtration, and an isoelectric point above 9.0. Atlantic cUNG is inhibited by the specific UNG inhibitor (Ugi) from the Bacillus subtilis bacteriophage (PBS2), and has a 2-fold higher activity for single-stranded DNA than for double-stranded DNA. cUNG has an optimum activity between pH 7.0-9.0 and 25-50 mM NaCl, and a temperature optimum of 41 degrees C. Cod UNG was compared with the recombinant human UNG (rhUNG), and was found to have slightly higher relative activity at low temperatures compared with their respective optimum temperatures. Cod UNG is also more pH- and temperature labile than rhUNG. At pH 10.0, the recombinant human UNG had 66% residual activity compared with only 0.4% for the Atlantic cUNG. At 50 degrees C, cUNG had a half-life of 0.5 min compared with 8 min for the rhUNG. These activity and stability experiments reveal cold-adapted features in cUNG.
authors: Department of Biotechnology, Institute of Medical Biology, Faculty of Medicine, University of Tromsø, Norway.
authors: O
authors: O
authors: Lanes
authors: P H
authors: PH
authors: Guddal
authors: D R
authors: DR
authors: Gjellesvik
authors: N P
authors: NP
authors: Willassen
mesh_terms: D000222:Adaptation, Physiological; D000818:Animals; D001267:Atlantic Ocean; D003080:Cold Temperature; D045647:DNA Glycosylases; D004795:Enzyme Stability; D005399:Fishes; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D009699:N-Glycosyl Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D012965:Sodium Chloride; D013379:Substrate Specificity; D013696:Temperature; D051981:Uracil-DNA Glycosidase
chemical_list: D012965:Sodium Chloride; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D051981:Uracil-DNA Glycosidase