title: An extracellular S1-type nuclease of marine fungus Penicillium melinii.
abstract: An extracellular nuclease was purified 165-fold with a specific activity of 41,250 U/mg poly(U) by chromatography with modified chitosan from the culture of marine fungus Penicillium melinii isolated from colonial ascidium collected near Shikotan Island, Sea of Okhotsk, at a depth of 123 m. The purified nuclease is a monomer with the molecular weight of 35 kDa. The enzyme exhibits maximum activity at pH 3.7 for DNA and RNA. The enzyme is stable until 75°C and in the pH range of 2.5-8.0. The enzyme endonucleolytically degrades ssDNA and RNA by 3'-5' mode to produce 5'-oligonucleotides and 5'-mononucleotides; however, it preferentially degrades poly(U). The enzyme can digest dsDNA in the presence of pregnancy-specific beta-1-glycoprotein-1. The nuclease acts on closed circular double-stranded DNA to produce opened circular DNA and then the linear form DNA by single-strand scission. DNA sequence encoding the marine fungus P. melinii endonuclease revealed homology to S1-type nucleases. The tight correlation found between the extracellular endonuclease activity and the rate of H³-thymidine uptake by actively growing P. melinii cells suggests that this nuclease is required for fulfilling the nucleotide pool of precursors of DNA biosynthesis during the transformation of hyphae into the aerial mycelium and conidia in stressful environmental conditions.
authors.affiliation: Pacific Institute of Bioorganic Chemistry, Department of Marine Biochemistry, Far Eastern Branch of Academy of Russia, Vladivostok, Russia. balaban@piboc.dvo.ru
authors.forename: Larissa A
authors.initials: LA
authors.lastname: Balabanova
authors.forename: Yury M
authors.initials: YM
authors.lastname: Gafurov
authors.forename: Mikhael V
authors.initials: MV
authors.lastname: Pivkin
authors.forename: Natalya A
authors.initials: NA
authors.lastname: Terentyeva
authors.forename: Galina N
authors.initials: GN
authors.lastname: Likhatskaya
authors.forename: Valery A
authors.initials: VA
authors.lastname: Rasskazov
mesh_terms: D000595:Amino Acid Sequence; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010407:Penicillium / Q000201:enzymology*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D005656:Fungal Proteins; D015719:Single-Strand Specific DNA and RNA Endonucleases