title: Hyperalkaline and thermostable phosphatase in Thermus thermophilus.
abstract: The phosphatases existing in the extreme thermophilic bacterium Thermus thermophilus have been studied. Utilizing ion exchange, hydrophobic, pseudoaffinity, and affinity chromatography, a number of distinct phosphatase activities were identified. At least four phosphatases, with optimum pH ranging between 5.0 and 11.5, were assayed with p-nitrophenylphosphate, and two with optimum pH between 7.0 and 11.0, with 32P-casein as substrate. The authors have focused on the hyperalkaline phosphatase and have tried to purify and characterize it. This hyperalkaline phosphatase reaches a maximal level at the stationary phase of the growth, and is co-purified with alkaline phosphatase with optimum pH of 10.2. The enzymes present a relative mol wt of 65 and 58 kDa, respectively, as judged by SDS-PAGE and Sephadex G-150 column, and possess similar properties, indicating that they are isoforms. These enzymes barely function in the presence of tartrate, and are inhibited by EDTA, pyrophosphate, and molybdate. Among the metals tested, Hg2+ appeared as the strongest inhibitor of the hyperalkaline phosphatase. The two enzymes are thermostable and, upon treatment at 90 degrees C for 10 min, 75% of their activity remains. The physiological role and function of these phosphatases need further investigation.
authors.affiliation: Department of Chemistry, Aristotle University of Thessaloniki, Greece.
authors.forename: A A
authors.initials: AA
authors.lastname: Pantazaki
authors.forename: A A
authors.initials: AA
authors.lastname: Karagiorgas
authors.forename: M
authors.initials: M
authors.lastname: Liakopoulou-Kyriakides
authors.forename: D A
authors.initials: DA
authors.lastname: Kyriakidis
mesh_terms: D000469:Alkaline Phosphatase / Q000302:isolation & purification / Q000378:metabolism; D001709:Biotechnology; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature; D016963:Thermus thermophilus / Q000201:enzymology*
chemical_list: D000469:Alkaline Phosphatase; D010749:Phosphoprotein Phosphatases; D010744:Phosphoric Monoester Hydrolases