Journal of Comparative Physiology B

, Volume 160, Issue 2, pp 201–206 | Cite as

Effect of anoxia on the kinetic properties of pyruvate kinase and phosphofructokinase, and on glycogen phosphorylase activity in marine worms and earth worms

  • Basile Michaelidis
  • Athanasios Papadopoulos
  • Isidoros Beis


The kinetic properties of PK and PFK were studied in aerobic versus 12-hours anoxic marine worms Hedistae(=Nereis) diversicolor and Diopatra neapolitana and earth worms Allolobophora calliginosa and Eisenia foetida. The total glycogen phosphorylase (a+b) activity and the percentage of active a form were also measured in the marine and earth worms under the same conditions. Anoxia exposure did not result in any significant changes of kinetic parameters of PK and total activities of glycogen phosphorylase from marine worms, but it altered the kinetic characteristics of PFK from H. diversicolor. Chromatographical studies showed that PK from both aerobic and anoxic marine worms is eluted from DEAE-cellulose as a single peak at 50 mM KCl. In contrast to marine worms, however, anoxia caused a marked change in kinetic properties of PK from both earth worms, resulting in a reduction of enzyme affinity for its substrate PEP. In addition, the enzyme existed in both earth worms in two distinct variants eluted from DEAE-cellulose column as peak I and peak II at 50 mM and 150 mM KCl, respectively. The ratio of enzyme units (peak I/peak II) was reduced significantly after 12 h of anoxia, indicating that these two peaks are interconvertible. Anoxia also caused a reduction of total glycogen phosphorylase activity in E. foetida and lowered the percentage of active a form of the enzyme by approximately 50% in both earth worms. Kinetic properties of PFK from both earth worms were not significantly affected by anoxia. However, their low Ka values for F-2,6-P2 imply that this effector may play an important role in PFK control in earth worms under anoxia.

Key words

Worms Enzymes Anoxia Metabolic regulation 











6-phosphofructo-1-kinase (E.C.


pyruvate kinase (E.C.


inorganic phosphate


phenyl methylsulfonyl fluoride


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  1. Famme P, Knudsen J, Hansen ES (1981) The effect of oxygen on the aerobic-anaerobic metabolism of the marine bivalve Mytilus edulis L. Mar Biol Lett 2: 345–351Google Scholar
  2. Gäde G (1983) Energy metabolism of arthropods and molluscs during environmental and functional anaerobiosis. J Exp Zool 228: 415–429Google Scholar
  3. Gruner B, Zebe E (1978) Studies on the anaerobic metabolism of earthworms. Comp Biochem Physiol 60B: 441–445Google Scholar
  4. Hofer WH, Allen LB, Kaeini MR, Harris BG (1982) Phosphofructokinase from Ascaris suum: The effect of phosphorylation on activity near-physiological conditions J Biol Chem 257: 3807–3810Google Scholar
  5. Hochachka PW, Somero GN (1984) Biochemical adaptations. Princeton University Press, Princenton New Jersey USAGoogle Scholar
  6. Holwerda DA, Veenhof PR, van Heugten HA, Zandee DI (1983) Regulation of mussel pyruvate kinase during anaerobiosis and in temperature acclimation by covalent modification. Mol Physiol 3: 225–234Google Scholar
  7. Hue L, Rider MH (1987) Role of fructose-2,6-bisphosphate in the control of glycolysis in mammalian tissues. Biochem J 245: 313–324Google Scholar
  8. Job D, Cochet C, Dhien A, Chambaz EM (1978) A rapid method for screening inhibitor effects: Determination of I50 and its standard deviation. Anal Biochem 84: 68–77Google Scholar
  9. Kamemoto ES, Mansour TE (1986) Phosphofructokinase in the liver fluke Fasciola hepatica: Purification and kinetic changes by phosphorylation. J Biol Chem 261: 4346–4351Google Scholar
  10. Livingstone DR, Zwaan A de (1983) Carbohydrate metabolism of gastropods In: Wilbour KM (ed) The Mollusca, vol 1. Academic Press, New York, pp 177–242Google Scholar
  11. Michaelidis B, Storey BK (1990a) Evidence for phosphorylation/dephosphorylation control of phosphofructokinase from organs of the anoxia-tolerant sea mussel Mytilus edulis L. J Exp Zool, in pressGoogle Scholar
  12. Michaelidis B, Storey BK (1990b) Phosphofructokinase from the anterior byssus retractor muscle of Mytilus edulis L: Modification of the enzyme in anoxia and by endogenous protein kinases. Int J Biochem, in pressGoogle Scholar
  13. Michaelidis B, Gaitanaki K, Beis I (1988) Modification of pyruvate kinase from the foot muscle of Patella caerulea L during anaerobiosis. J Exp Zool 248: 264–271Google Scholar
  14. Papadopulos A, Michaelidis B, Beis IS (1989) Pyruvate kinase from the earth worm Allolobophora calliginosa: Modification of the enzyme during anaerobiosis possibly by phosphorylation/dephosphorylation. Can J Zool, in pressGoogle Scholar
  15. Plaxton WC, Storey KB (1984a) Purification and properties of aerobic and anoxic forms of pyruvate kinase from red muscle tissue of the channelled whelk, Busycotypus canaliculatum. Eur J Biochem 143: 257–265Google Scholar
  16. Plaxton WC, Storey KB (1984b) Phosphorylation in vivo of red muscle pyruvate kinase from the channeled whelk, Busycotypus canaliculatum, in response to anoxic stress. Eur J Biochem 143: 267–272Google Scholar
  17. Schaftingen E van (1984) Fructose-2,6-bisphosphate. In: Bergmeyer HU (ed) Methods of enzymatic analysis, Vol VI. Verlag Chemie, Weinheim, pp 335–341Google Scholar
  18. Shick JM, de Zwaan A, de Bont AMT (1983) Anoxic metabolic rate in the Mytilus edulis L estimated by simultaneous direct calorimetry and biochemical analysis. Physiol Zool 56: 56–63Google Scholar
  19. Schöttler U (1978a) Investigations of the anaerobic metabolism of the polychaete Nereis diversicolor M. J Comp Physiol 125: 185–189Google Scholar
  20. Schöttler U (1978b) The influence of anaerobiosis on the levels of adenosine nucleotides and some glycolytic metabolites in Tubifex sp. (Annelida, Oligochaeta). Comp Biochem Physiol 61 B: 29–32Google Scholar
  21. Schöttler U, Wienhausen G, Zebe E (1983) The mode of energy production in the lugworm Arenicola marina at different oxygen concentrations. J Comp Physiol 149: 547–555Google Scholar
  22. Storey KB (1984) Phosphofructokinase from foot muscle of the whelk, Busycotypus canaliculatum: Evidence for covalent modification of the enzyme during anaerobiosis. Arch Biochem Biophys 235: 665–672Google Scholar
  23. Storey KB (1988) Mechanisms of glycolytic control during facultative anaerobiosis in a marine mollusc: tissue-specific analysis of glycogen phosphorylase and fructose-2,6-biphosphate. Can J Zool 66: 1767–1771Google Scholar
  24. Zwaan A de (1983) Carbohydrate metabolism in bivalves. In: Wilbour KM (ed) The mollusca, vol 1. Academic Press, New York, pp 137–175Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Basile Michaelidis
    • 1
  • Athanasios Papadopoulos
    • 1
  • Isidoros Beis
    • 1
  1. 1.Laboratory of Animal Physiology, Department of Zoology, Science SchoolUniversity of ThessalonikiThessalonokiGreece

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