Protein from the soluble fraction of Bacterium matruchotii cells propagated in a medium containing no added inorganic phosphate was fractionated by gel permeation chromatography. The bulk of phosphatase activity assayed at pH 8.0 was found in fractions equivalent to a molecular weight of 6 × 105 daltons. Substrate saturation kinetics indicate at Km of 0.75 mM for p-nitrophenylphosphate. Activity was stimulated more than twofold by Zn++ at 1 mM, but was significantly reduced by EDTA, Ca++ and inorganic phosphate. The enzyme(s) shows negligible activity at pH below 6.0 and has a narrow optimum between 7.5 and 8.5.
Get full access to this article
View all access options for this article.
References
1.
Howell, A., Jr.; and Fitzgerald, R.J.: The production of acid phosphatases by certain species of Actinomyces, J. Bacteriol., 66 :437-442,1953.
2.
Ennever, J.J. ; and Warner, B.W.: Phosphatase and the Oral Actinomyces, J. Dent. Res., 31:25-26, 1952.
3.
Garen, A.; and Levinthal, C.: A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase, Biochem. Biophys. Acta, 38 :470-483, 1960.
4.
Halford, S.E. ; Bennet, N.G.; Tren-Tham, D.R.; and Gutfreund , H.: A substrate-induced conformation change in the reaction of alkaline phosphatase from Escherichia coli, Biochem. J., 114:243-251, 1969.
5.
Plocke, D.J. ; Levinkal, C.; and Vallee, B.L.: Alkaline phosphatase of Escherichia coli: A zinc metalloenzyme, Biochemistry, 1 :373-378, 1962.
6.
Heppel, L.A. ; Harkenss, D.R.; and Hilmore, R.J.: A study of the substrate specificity and other properties of the alkaline phosphatase of Escherichia coli, J. Biol. Chem., 237 :841-846, 1962.
7.
Gron, P.: The state of calcium and inorganic orthophosphate in human saliva, Arch. Oral Biol., 18:1365-1378, 1973 .
8.
Ennever, J.; Vogel, J.J.; and Streck-Fuss, J.L. : Synthetic medium for calcification of Bacterionema matruchotii, J. Dent. Res., 50:1327-1330, 1971.
