Applied Microbiology and Biotechnology

, Volume 42, Issue 2–3, pp 263–269 | Cite as

Effect of ferric and cupric ions on the inactivation rate of dextransucrase

  • R. W. Lencki
  • M. Delaire
  • A. Tecante
  • L. Choplin
Original Paper
  • 35 Downloads

Abstract

When ferric ion was added to solutions of the enzyme dextransucrase, first-order followed by second-order inactivation behavior was observed. The initial rapid activity loss was attributed to a ferric ion interacting with the thiol group of the native monomer to form a less active enzyme-ion complex; the second inactivation stage involved enzyme-ion complex aggregation and disulfide cross-link formation. In contrast, Cu2+ ion inactivation demonstrated simple first-order kinetics. As with Fe3+, Cu2+ ions can form complexes with enzyme thiol groups. However, unlike ferric ions, cupric ions can also strongly interact with the imidazole ring of histidine. Since the dextransucrase active site contains two key histidines, imidazole-cupric-ion interactions could potentially inhibit enzymatic activity. Thus, it was hypothesized that first-order Cu2+ inactivation kinetics involved the adsorption of this ion to the enzyme's activity site. The addition of a reducing agent such as dithiothreitol can inhibit the second enzyme aggregation stage by breaking disulfide cross-links but cannot restrict the initial formation of metal-enzyme complexes.

Keywords

Histidine Thiol Group Imidazole Ring Inactivation Rate Inactivation Kinetic 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Agarwal PK (1985) Heterogeneous denaturation of enzymes: a distributed activation energy model with nonuniform activities. Biotechnol Bioeng 27:1554–1563Google Scholar
  2. Baldwin RL (1975) Intermediates in protein folding reactions and the mechanisms of protein folding. Annu Rev Biochem 44:453–475Google Scholar
  3. Boyer PD (1959) Sulfhydryl and disulfide groups of enzymes. In: Boyer PD, Lardy H, Myrbäck K (eds) The enzymes, vol 1. Academic Press, New York, pp 511–588Google Scholar
  4. Breslow E (1964) Comparison of cupric ion-binding sites in myoglobin derivatives and serum albumin. J Biol Chem 239:3252–3259Google Scholar
  5. Caughey WS, Wallace WJ, Volpe JA, Yoshikawa S (1976) Cytochrome oxidase. In: Boyer, PD (ed) The enzymes, vol 13. Academic Press, New York, pp 299–344Google Scholar
  6. Cleland WW (1964) Dithiothreitol, a new protective reagent for SH groups. Biochemistry 3:480–482Google Scholar
  7. Dixon M, Webb EC (1979) Enzymes, 3rd edn. Academic Press, New York, pp 271–331Google Scholar
  8. Downes JM, Whelan J, Bosnich B (1981) Biological analogues. Spectroscopic characteristics of mercapto- and disulfide-copper(II) coordination in relation to type I proteins. Inorg Chem 20:1081–1086Google Scholar
  9. Ehrenberg L, Harms-Ringdahl M, Fedorcsák I, Granath F (1989) Kinetics of the copper- and iron-catalyzed oxidation of cysteine by dioxygen. Acta Chem Scand 43:177–187Google Scholar
  10. Fishbean WN, Nagarajan K (1971) Urease catalysis and structure VII. Factors involved in urease polymerization and its kinetic pattern. Arch Biochem Biophys 144:700–714Google Scholar
  11. Fu D, Robyt JF (1988) Essential histidine residues in dextransucrase: chemical modification by diethyl pyrocarbonate and dye photooxidation. Carbohydrate Res 183:97–109Google Scholar
  12. Hamed MY, Silver J (1983) Studies of the reaction of ferric ion with glutathione and some related thiols. Part II. Complex formation in the pH range of three to seven. Inorg Chim Acta 78:1–11Google Scholar
  13. Hanaki A, Kamide H (1973) Manometric study of the copper-catalyzed oxidation of cysteine. Chem Pharm Bull (Tokyo) 19:1006–1010Google Scholar
  14. Kaboli H, Reilly PJ (1980) Immobilization and properties ofLeuconostoc mesenteroides dextransucrase. Biotechnol Bioeng 22:1055–1069Google Scholar
  15. Kobayashi M, Matsuda K (1976) Purification and properties of the extracellular dextransucrase fromLeuconostoc mesenteroides NRRL B-1299. J Biochem (Tokyo) 79:1301–1308Google Scholar
  16. Lencki RW, Arul J, Neufeld RJ, (1992a) Effect of subunit dissociation, denaturation aggregation and coagulation on enzyme inactivation kinetics. I. First-order behavior. Biotechnol Bioeng 40:1421–1426Google Scholar
  17. Lencki RW, Arul J, Neufeld RJ (1992b) Effect of subunit dissociation, denaturation aggregation and coagulation on enzyme inactivation kinetics. II. Biphasic and grace period behavior. Biotechnol Bioeng 40:1427–1434Google Scholar
  18. Lencki RW, Tecante A, Choplin L (1993) Effect of shear on the inactivation kinetics of the enzyme dextransucrase. Biotechnol Bioeng 42:1061–1067Google Scholar
  19. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428Google Scholar
  20. Paul F, Auriol D, Oriol E, Monsan P (1984) Production and purification of dextransucrase fromLeuconostoc mesenteroides, NRRL B512(F). Enzyme Eng 434:267–270Google Scholar
  21. Porath J, Carlsson J, Olsson I, Belfrage G (1975) Metal chelate affinity chromatography, a new approach to protein fractionation. Nature 258:598–599Google Scholar
  22. Reiner JM (1969) Behaviour of enzyme systems, 2nd ed. van Nostrand Reinhold, New York, pp 293–303Google Scholar
  23. Robyt JE, Walseth TE (1979) Production, purification and properties of dextransucrase fromLeuconostoc mesenteroides NRRLB-512F. Carbohydr Res 68:95–111Google Scholar
  24. Sadana A, Henley JP (1987) Single-step unimolecular non-firstorder enzyme deactivation kinetics. Biotechnol Bioeng 30:717–723Google Scholar
  25. Taylor JE, Yan JF, Wang J-H (1966) The iron(III)-catalyzed oxidation of cysteine by molecular oxygen in the aqueous phase. An example of a two-thirds-order reaction. J Am Chem Soc 88:1663–1667Google Scholar
  26. Tirrell M, Middleman S (1975) Shear modification of enzyme kinetics. Biotechnol Bioeng 17:299–303Google Scholar
  27. Tirrell M, Middleman S (1978) Shear deformation effects in enzyme catalysis. Biophys J 23:121–128Google Scholar
  28. Torchinskii IM (1981) Sulfur in proteins. Pergamon Press, Oxford, pp 123–135Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • R. W. Lencki
    • 1
  • M. Delaire
    • 1
  • A. Tecante
    • 1
  • L. Choplin
    • 2
  1. 1.Department of Chemical EngineeringUniversité LavalSainte-FoyQuébecCanada
  2. 2.Department of Food ScienceUniversity of GuelphGuelphCanada

Personalised recommendations