Summary
Partially purified fructose diphosphatase from the obligate chemolithotroph,Thiobacillus neapolitanus has been characterized, and some of its regulatory properties described. The enzyme had a high effinity for its substrate, but was inhibited by substrate at concentrations above 1 mM. The enzyme had an absolute requirement for a divalent cation. In the absence of EDTA there was a single pH optimum in the alkaline range between 8.5 and 9.5; in the presence of EDTA there was considerable was activity at both neutral and alkaline pH. This diphosphatase was inhibited by AMP at 10−4 M or greater-, the lower the pH, the greater the AMP inhibition. Treatment of the enzyme with 5×10−5 Mpara hydroxy mercuribenzoate allowed retention of full catalytic activity while abolishing considerable AMP inhibition. Exposure of the enzyme to several concentrations of urea had no effect on the AMP inhibition. Homocystine (0.06 mM) and coenzyme A (0.1 mM) had no effect. At 1 mM, PEP caused 60% inhibition, 2, 3-diphosphoglyceric acid produced 26% inhibition, and pyruvate had no effect.
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References
Ballard, R. W., macElroy, R. D.: Phosphoenolpyruvate, a new inhibitor of phosphoribulokinase inPseudonomas facilis. Biochem. biophys. Res. Commun.44, 614–618 (1971).
Buchanan, B. B., Arnon, D. I.: Ferredoxins: Chemistry and function in photosynthesis, nitrogen fixation and fermentative metabolism. In: Advances in enzymology, Vol. 33, pp. 119–176. F. F. Nord, Ed. New York: Interscience Publishers 1971.
Cornish, A. S., Johnson, E. J.: Regulation of pyruvate kinase fromThiobacillus neapolitanus. Arch. Biochem. Biophys.142, 584–590 (1971).
Johnson, E. J., Abraham, S.: Enzymes of intermediary carbohydrate metabolism in the obligate autotrophsThiobacillus thioparus andThiobacillus neapolitanus. J. Bact.100, 962–968 (1969).
Kirtley, M. E., Dix, J. C.: Activation of fructose diphosphatase by manganese, magnesium and cobalt. Arch. Biochem. Biophys.147, 647–652 (1971).
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurements with the Folin phenol reagent. J. biol. Chem.193, 265–275 (1951)
Mokrasch, L. C., McGilvery, R. W.: Purification and properties of fructose-1,6-diphosphatase. J. biol. Chem.221, 909–917 (1956)
Mukkada, A. J., Bell, E. J.: Fructose-1,6-diphosphatase ofAcinetobacter; Inhibition by ATP and citrate. Biochem. biophys. Res. Commun.37, 340–346 (1969)
Mukkada, A. J., Bell, E. J.: Partial purification and properties of the fructose-1,6-diphosphatase fromAcinetobacter lwoffi. Arch. Biochem. Biophys.142, 22–31 (1971)
Nakashima, K., Pontremoli, S., Horecker, B. L.: Activation of rabbit liver fructose diphosphatase by coenzyme A and acyl carrier protein. Proc. nat. Acad. Sci. (Wash.)64, 947–951 (1969)
Pontremoli, S., Horecker, B. L.: Fructose-1,6-diphosphatase from rabbit liver. In: Current topics in cellular regulation, Vol. 2, pp. 173–199, B. L. Horecker, E. R. Stadtman, Eds. New York: Academic Press 1970
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Johnson, E.J., MacElroy, R.D. Regulation in the chemolithotrophthiobacillus neapolitanus: Fructose-1,6-diphosphatase. Archiv. Mikrobiol. 93, 23–28 (1973). https://doi.org/10.1007/BF00666078
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DOI: https://doi.org/10.1007/BF00666078