Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1986 Oct;82(2):462–467. doi: 10.1104/pp.82.2.462

Isolation and Characterization of Two Enzymes Capable of Hydrolyzing Fructose-1,6-Bisphosphatase from the Lichen Peltigera rufescens1

Doug Brown 1, Kenneth A Kershaw 1
PMCID: PMC1056141  PMID: 16665052

Abstract

Two enzymes capable of hydrolyzing fructose-1,6-bisphosphate (FBP) have been isolated from the foliose lichen Peltigera rufescens (Weis) Mudd. These enzymes can be separated using Sephadex G-100 and DEAE Sephacel chromatography. One enzyme has a pH optimum of 6.5, and a substrate affinity of 228 micromolar FBP. This enzyme does not require MgCl2 for activity, and is inhibited by AMP. The second enzyme has a pH optimum of 9.0, with no activity below pH 7.5. This enzyme responds sigmoidally to Mg2+, with half-saturation concentration of 2.0 millimolar MgCl2, and demonstrates hyperbolic kinetics for FBP (Km = 39 micromolar). This enzyme is activated by 20 millimolar dithiothreitol, is inhibited by AMP, but is not affected by fructose-2-6-bisphosphate. It is hypothesized that the latter enzyme is involved in the photosynthetic process, while the former enzyme is a nonspecific acid phosphatase.

Full text

PDF
467

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bishop R. H. Regulatory characteristics of a fructose bisphosphatase from the blue-green bacterium Anacystis nidulans. Arch Biochem Biophys. 1979 Aug;196(1):295–300. doi: 10.1016/0003-9861(79)90579-4. [DOI] [PubMed] [Google Scholar]
  2. Brown D., Kershaw K. A. Seasonal Changes in the Kinetic Parameters of a Photosynthetic Fructose-1,6-Bisphosphatase Isolated from Peltigera rufescens. Plant Physiol. 1986 Oct;82(2):457–461. doi: 10.1104/pp.82.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buchanan B. B., Schürmann P., Kalberer P. P. Ferredoxin-activated fructose diphosphatase of spinach chloroplasts. Resolution of the system, properties of the alkaline fructose diphosphatase component, and physiological significance of the ferredoxin-linked activation. J Biol Chem. 1971 Oct 10;246(19):5952–5959. [PubMed] [Google Scholar]
  4. Charles S. A., Halliwell B. Properties of freshly purified and thiol-treated spinach chloroplast fructose bisphosphatase. Biochem J. 1980 Mar 1;185(3):689–693. doi: 10.1042/bj1850689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cséke C., Weeden N. F., Buchanan B. B., Uyeda K. A special fructose bisphosphate functions as a cytoplasmic regulatory metabolite in green leaves. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4322–4326. doi: 10.1073/pnas.79.14.4322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Funayama S., Molano J., Gancedo C. Purification and properties of a D-fructose 1,6-bisphosphatase from Saccharomyces cerevisiae. Arch Biochem Biophys. 1979 Oct 1;197(1):170–177. doi: 10.1016/0003-9861(79)90233-9. [DOI] [PubMed] [Google Scholar]
  7. Gerbling K. P., Steup M., Latzko E. Fructose-1,6-bisphosphatase from Synechococcus leopoliensis. Substrate-dependent dimer-tetramer interconversion. Eur J Biochem. 1985 Feb 15;147(1):207–215. doi: 10.1111/j.1432-1033.1985.tb08738.x. [DOI] [PubMed] [Google Scholar]
  8. Herzog B., Stitt M., Heldt H. W. Control of Photosynthetic Sucrose Synthesis by Fructose 2,6-Bisphosphate : III. Properties of the Cytosolic Fructose 1,6-Bisphosphatase. Plant Physiol. 1984 Jul;75(3):561–565. doi: 10.1104/pp.75.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Latzko E., Zimmermann G., Feller U. Evidence for a hexosediphosphatase from the cytoplasm of spinach leaves. Hoppe Seylers Z Physiol Chem. 1974 Mar;355(3):321–326. doi: 10.1515/bchm2.1974.355.1.321. [DOI] [PubMed] [Google Scholar]
  10. Pradel J., Soulié J. M., Buc J., Meunier J. C., Ricard J. On the activation of fructose-1,6-bisphosphatase of spinach chloroplasts and the regulation of the Calvin cycle. Eur J Biochem. 1981 Jan;113(3):507–511. doi: 10.1111/j.1432-1033.1981.tb05092.x. [DOI] [PubMed] [Google Scholar]
  11. Preiss J., Biggs M. L., Greenberg E. The effect of magnesium ion concentration on the pH optimum of the spinach leaf alkaline fructose diphosphatase. J Biol Chem. 1967 May 10;242(9):2292–2294. [PubMed] [Google Scholar]
  12. RACKER E., SCHROEDER E. A. The reductive pentose phosphate cycle. II. Specific C-1 phosphatases for fructose 1,6-diphosphate and sedoheptulose 1,7-diphosphate. Arch Biochem Biophys. 1958 Apr;74(2):326–344. doi: 10.1016/0003-9861(58)90004-3. [DOI] [PubMed] [Google Scholar]
  13. Rosa L., Whatley F. R. Conditions Required for the Rapid Activation In Vitro of the Chloroplast Fructose-1,6-bisphosphatase. Plant Physiol. 1984 May;75(1):131–137. doi: 10.1104/pp.75.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Stitt M., Herzog B., Heldt H. W. Control of Photosynthetic Sucrose Synthesis by Fructose 2,6-Bisphosphate : V. Modulation of the Spinach Leaf Cytosolic Fructose 1,6-Bisphosphatase Activity in Vitro by Substrate, Products, pH, Magnesium, Fructose 2,6-Bisphosphate, Adenosine Monophosphate, and Dihydroxyacetone Phosphate. Plant Physiol. 1985 Nov;79(3):590–598. doi: 10.1104/pp.79.3.590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Udvardy J., Godeh M. M., Farkas G. L. Regulatory properties of a fructose 1,6-bisphosphatase from the cyanobacterium Anacystis nidulans. J Bacteriol. 1982 Jul;151(1):203–208. doi: 10.1128/jb.151.1.203-208.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Waley S. G. An easy method for the determination of initial rates. Biochem J. 1981 Mar 1;193(3):1009–1012. doi: 10.1042/bj1931009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Zimmermann G., Kelly G. J., Latzko E. Efficient purification and molecular properties of spinach chloroplast fructose 1,6-bisphosphatase. Eur J Biochem. 1976 Nov 15;70(2):361–367. doi: 10.1111/j.1432-1033.1976.tb11025.x. [DOI] [PubMed] [Google Scholar]
  18. Zimmermann G., Kelly G. J., Latzko E. Purification and properties of spinach leaf cytoplasmic fructose-1,6-bisphosphatase. J Biol Chem. 1978 Sep 10;253(17):5952–5956. [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES