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Isolation and properties of two DNA-polymerases from Bacillus stearothermophilus cells

Trofimenko, A.F.; Gaziev, A.I.

Biokhimiia 46(4): 681-690

1981

ISSN/ISBN: 0320-9725
PMID: 7284483
Document Number: 175073
Two DNA-polymerases (1 and 2) were isolated from the cells of Bacillus stearothermophilus and their properties were investigated. The isolation procedure included separation of nucleic acids on DEAE-cellulose, affinity chromatography on DNA-agarose and chromatography on DEAE-Sephadex. DNA-polymerase 1 was purified 150-fold; its molecular weight and pH optimum are 135 000 and 7.3, respectively. DNA-polymerase 2 was purified 1000-fold; its molecular weight and pH optimum are 95 000 and 9.0, respectively. Both DNA polymerases differ considerably in their affinities for DNA-agarose and DEAE-Sephadex. Purification of DNA-polymerase 1 resulted in a loss of the enzyme thermal stability and its conversion into a labile enzyme with the temperature optimum at 40 degrees. DNA-polymerase 2 retained its high thermal stability after purification. The DNA activated by DNAase 1 was found to be the most effective primer matrix for the both enzymes. In contrast to DNA-polymerase 2, DNA-polymerase 1 is sensitive to the effects of SH-group blockers and high salt concentrations and is activated 1.5 -- 2-fold by 10% ethanol.

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Sokolov, N.N.; Votrin, I.I.; Fitzner, A.B.; Anikejcheva, N.V. 1978: Isolation and some properties of restriction endonuclease from Bacillus amyloliquefaciens Biokhimiia 43(5): 865-871
Koltukova, N.V.; Bondarchuk, A.A.; Zakharova, I.I. 1982: Isolation, purification and properties of Bacillus mesentericus proteinases Mikrobiolohichnyi Zhurnal 44(3): 12-15
Hennig, W.A.; Stern, H.S.; Lanza, R.J.; Murthy, Y.S. 1973: Use of Bacillus stearothermophilus as a biological indicator Bulletin of the Parenteral Drug Association 27(1): 30-37
Penna, T.C.; Machoshvili, I.A.; Taqueda, M.E.; Ferraz, C.A. 1998: Bacillus stearothermophilus sporulation response to different composition media PDA Journal of Pharmaceutical Science and Technology 52(5): 198-208
Ouderkirk, L.A. 1977: Detection of residual penicillins in milk by using a Bacillus stearothermophilus disk assay Journal - Association of Official Analytical Chemists 60(5): 1116-1118
Amelunxen, R.E.; Sauvan, R.L.; Mira, O.J. 1974: Thermostable glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus: II. Kinetic and immunokinetic analysis Physiological Chemistry and Physics 6(6): 515-526
Koyama, T.; Obata, S.; Osabe, M.; Saito, K.; Takeshita, A.; Nishino, T.; Ogura, K. 1994: Thermostable farnesyl diphosphate synthase of Bacillus stearothermophilus: crystallization and site-directed mutagenesis Acta Biochimica Polonica 41(3): 281-292
Sakanian, V.A.; Legreĭn, K.; Charlier, D.; Kochikian, A.V. 1993: N-acetylglutamate-5-phosphotransferase of the thermophilic bacterium Bacillus stearothermophilus: nucleotide sequence of the gene and enzyme characterization Genetika 29(4): 556-570
Oleĭnikova, G.K.; Shevchenko, L.S.; Kuznetsova, T.A.; Mikhaĭlov, V.V. 1995: A novel fungicide of the iturin group, obtained from a marine isolate of Bacillus subtilis. Isolation, physico-chemical and biochemical properties, identification Antibiotiki i Khimioterapiia 40(2): 19-21
Philippopoulos, M.; Xiang, Y.; Lim, C. 1995: Identifying the mechanism of protein loop closure: a molecular dynamics simulation of the Bacillus stearothermophilus LDH loop in solution Protein Engineering 8(6): 565-573
Spicher, G.; Peters, J. 1995: Microbial resistance to formaldehyde. III> Dependence of the microbial effect on Staphylococcus aureus, Enterococcus faecium and spores of Bacillus stearothermophilus on temperature Zentralblatt für Hygiene und Umweltmedizin 196(6): 545-561
Bezuglyĭ, S.V.; Skripal', I.G.; Babichev, V.V. 1992: The chromatographic properties of the DNA-dependent DNA polymerases from Acholeplasma laidlawii PG-8 Mikrobiolohichnyi Zhurnal 54(1): 51-57
Shirokova, E.A.; Shipitsyn, A.V.; Kuznetsova, E.V.; Viktorova, L.S.; Kraevskiĭ, A.A. 1993: 5'-S-nucleoside triphosphates. Synthesis and substrate properties in tests with DNA polymerases Bioorganicheskaia Khimiia 19(12): 1226-1233
Kaiho, T.; Sekiya, S.; Shirotake, S.; Takamizawa, H. 1984: Isolation and properties of methotrexate-resistant choriocarcinoma cells in vitro Nihon Sanka Fujinka Gakkai Zasshi 36(12): 2517-2524
Goncharov, E.K.; Suchkov, I.I.; Mishan'kin, B.N. 1984: Isolation, purification and physicochemical properties of pesticin i from cells of Yersinia pestis strain EV Zhurnal Mikrobiologii Epidemiologii i Immunobiologii 6: 41-44
Nersesova, L.S.; Ashmarin, I.P.; Lyzlova, S.N. 1976: Creatine kinase of the peritoneal macrophages: isolation, its connection with the functional activity of the cells and certain properties Zhurnal Eksperimental'noi i Klinicheskoi Meditsiny 16(1): 59-65
Akimkina, T.V.; Hosovskaia, E.A.; Kostrov, S.V. 1992: Cloning and gene expression of Bacillus cereus neutral proteinase in Bacillus subtilis cells Molekuliarnaia Biologiia 26(2): 418-423
Bashkirov, V.I.; Vasil'ev, V.V.; Prozorov, A.A. 1988: Cloning in cells of Bacillus subtilis and study of the characteristics of promoter regions of DNA of Bacillus mesentericus and Phage SPO2 Molekuliarnaia Genetika Mikrobiologiia i Virusologiia 1: 13-20
Sekeris, C.E.; Economidis, J.V.; Hatzoglou, M. 1982: Homoribonucleotide polymerases of eucaryotic cells Progress in Clinical and Biological Research 102 Pt C: 139-152