Journal of Applied Phycology

, Volume 20, Issue 3, pp 271–278 | Cite as

Vanadium-dependent bromoperoxidases from Gracilaria algae

  • Tuangporn SuthiphongchaiEmail author
  • Patcharee Boonsiri
  • Bhinyo Panijpan


Red algae from the Gulf of Thailand were examined for haloperoxidatic activity. Six species, Gracilaria changii, G. edulis, G. firma, G. fisheri, G. salicornia, and G. tenuistipitata, showed bromoperoxidatic activity. Duplicate polyacrylamide electrophoretic gels showed enzyme activity patterns developed by phenol red staining for bromoperoxidatic activity and by 3,3′-diaminobenzidine staining for peroxidatic activity. All algae gave isoenzymic bromoperoxidatic activity bands and peroxidatic activity bands, but there were peroxidatic and bromoperoxidatic activity bands that did not correspond. The bromoperoxidatic activity of the crude enzyme extracts as well as previously dialyzed enzyme solutions was enhanced significantly by incubation with vanadium pentoxide. The three purified bromoperoxidases from G. fisheri contained vanadium, and their relative activities corresponded to the ratio of vanadium to enzyme. In addition, they were not inhibited by H2O2. These data confirm that the enzymes are vanadium bromoperoxidases.


Bromoperoxidase staining Red algae V-BPO 











Low-molecular-weight bromoperoxidase from G. fisheri


High-molecular-weight bromoperoxidase from G. fisheri




Unit of enzyme activity (μmole MCD decrease.min−1)


Vanadium-reactivated apo-bromoperoxidase


Vanadium bromoperoxidase



This work was supported by a grant from the National Research Council of Thailand. We thank Prof. Dr. Khanjanapaj Lewmanomont Faculty of Fisheries, Kasetsart University, for her assistance in identification of algal samples.


  1. Ahern TJ, Allan GG, Medcalf DG (1980) New bromoperoxidases of marine origin: partial purification and characterization. Biochim Biophys Acta 616:329–339PubMedGoogle Scholar
  2. Alaee M, Arias P, Sjodin A, Bergman A (2003) An overview of commercially used brominated flame retardants, their applications, their use patterns in different countries/regions and possible modes of release. Environ Int 29:683–689PubMedCrossRefGoogle Scholar
  3. Almeida M, Filipe S, Humanes M, Maia MF, Melo R, Severino N, da Silva JA, Frausto da Silva JJ, Wever R (2001) Vanadium haloperoxidases from brown algae of the Laminariaceae family. Phytochemistry 57:633–642PubMedCrossRefGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  5. Butler A, Carter-Franklin JN (2004) The role of vanadium bromoperoxidase in the biosynthesis of halogenated marine natural products. Nat Prod Rep 21:180–188PubMedCrossRefGoogle Scholar
  6. Cardozo KH, Guaratini T, Barros MP, Falcao VR, Tonon AP, Lopes NP, Campos S, Torres MA, Souza AO, Colepicolo P, Pinto E (2007) Metabolites from algae with economical impact. Comp Biochem Physiol C Toxicol Pharmacol 146:60–78PubMedCrossRefGoogle Scholar
  7. Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann N Y Acad Sci 121:404–427PubMedCrossRefGoogle Scholar
  8. de Boer E, Van Kooyk Y, Tromp MGM, Plat H, Wever R (1986) Bromoperoxidase from Ascophyllum nodosum: a novel class of enzymes containing vanadium as a prosthetic group? Biochim Biophys Acta 869:48–53Google Scholar
  9. de Boer E, Plat H, Tromp MGM, Wever R, Franssen MCR, van der Plas HC, Meije EM, Schoemaker HE (1987) Vanadium containing bromoperoxidase: an example of an oxidoreductase with high operational stability in aqueous and organic media. Biotechnol Bioeng 30:607–610CrossRefGoogle Scholar
  10. Everett RR, Kanofsky JR, Butler A (1990) Mechanistic investigations of the novel non-heme vanadium bromoperoxidases. Evidence for singlet oxygen production. J Biol Chem 265:4908–4914PubMedGoogle Scholar
  11. Fenical W (1975) Halogenation in the Rhodophyta. A review. J Phycol 11:245–259Google Scholar
  12. Greenwood NN, Earnshaw A (1984) Chemistry of the elements. Pergamon Press, OxfordGoogle Scholar
  13. Hager LP, Morris DR, Brown FS, Eberwein H (1966) Chloroperoxidase. II. Utilization of halogen anions. J Biol Chem 241:1769–1777PubMedGoogle Scholar
  14. Itoh N, Hasan AK, Izumi Y, Yamada H (1988) Substrate specificity, regiospecificity and stereospecificity of halogenation reactions catalyzed by non-heme-type bromoperoxidase of Corallina pilulifera. Eur J Biochem 172:477–484PubMedCrossRefGoogle Scholar
  15. Itoh N, Sasaki H, Ohsawa N, Shibata MS, Miura J (1996) Bromoperoxidase in Corallina pilulifera is regulated by its vanadate content. Phytochemistry 42:277–281CrossRefGoogle Scholar
  16. Kongkiattikajorn J, Pongdam S (2006) Vanadium haloperoxidase from the red alga Gracilaria fisheri. Sci Asia 32:25–30CrossRefGoogle Scholar
  17. Krenn BE, Plat H, Wever R (1987) The bromoperoxidase from the red alga Ceramium rubrum also contains vanadium as a prosthetic group. Biochim Biophys Acta 912:287–291Google Scholar
  18. Krenn BE, Izumi Y, Yamada H, Wever R (1989) A comparison of different (vanadium) bromoperoxidases; the bromoperoxidase from Corallina pilulifera is also a vanadium enzyme. Biochim Biophys Acta 998:63–68Google Scholar
  19. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  20. Lewmanomont K (1993) The species of Gracilaria from Thailand. In: Abbott IA (ed) Taxonomy of economic seaweeds, with reference to some Pacific species. Vol IV. California Sea Grant College, La Jolla, CA, pp 135–148Google Scholar
  21. Neidleman SL (1975) Microbial halogenation. CRC Crit Rev Microbiol 3:333–358PubMedCrossRefGoogle Scholar
  22. Neidleman SL, Geigert J (1986) Biohalogenation: principle, basic, roles and applications. Ellis Horwood, ChichesterGoogle Scholar
  23. Ohsawa N, Ogata Y, Okada N, Itoh N (2001) Physiological function of bromoperoxidase in the red marine alga, Corallina pilulifera: production of bromoform as an allelochemical and the simultaneous elimination of hydrogen peroxide. Phytochemistry 58:683–692PubMedCrossRefGoogle Scholar
  24. Olsen RL, Little C (1979) The peroxidase activity of rat uterus. Eur J Biochem 101:333–339PubMedCrossRefGoogle Scholar
  25. Pedersen M (1976) A brominating and hydroxylating peroxidase from the red alga Cystoclonium purpureum. Physiol Plant 37:6–11CrossRefGoogle Scholar
  26. Pereira RC, Da Gama BA, Teixeira VL, Yoneshigue-Valentin Y (2003) Ecological roles of natural products of the Brazilian red seaweed Laurencia obtusa. Braz J Biol 63:665–672PubMedCrossRefGoogle Scholar
  27. Polzin JJ, Rorrer GL, Cheney DP (2003) Metabolic flux analysis of halogenated monoterpene biosynthesis in microplantlets of the macrophytic red alga Ochtodes secundiramea. Biomol Eng 20:205–215PubMedCrossRefGoogle Scholar
  28. Rehder D (1991) The bioinorganic chemistry of vanadium. Angew Chem Int Ed 30:148–167Google Scholar
  29. Soedjak HS, Butler A (1991) Mechanism of dioxygen formation catalyzed by vanadium bromoperoxidase from Macrocystis pyrifera and Fucus distichus: steady state kinetic analysis and comparison to the mechanism of V-BrPO from Ascophyllum nodosum. Biochim Biophys Acta 1079:1–7PubMedGoogle Scholar
  30. Suthiphongchai T, Boonsiri P, Lewmanomont K, Panijpan B (1994) Purification of bromoperoxidases from Gracilaria fisheri. Paper presented at 11th FAOBMB Symposium: Biopolymers and Bioproducts: Structure, Function and Applications, Bangkok, Thailand, 15–18 November 1994Google Scholar
  31. Vilter H (1984) Peroxidases from phaeophyceae: a vanadium(V)-dependent peroxidase from Ascophyllum nodosum. Phytochemistry 23:1387–1390CrossRefGoogle Scholar
  32. Vimokesant S, Kunjara S, Rungruangsak K, Nakornchai S, Panijpan B (1982) Beriberi caused by antithiamin factors in food and its prevention. Ann N Y Acad Sci 378:123–136PubMedCrossRefGoogle Scholar
  33. Wever R, Plat H, de Boer E (1985) Isolation procedure and some properties of the bromoperoxidase from the seaweed Ascophyllum nodosum. Biochim Biophys Acta 830:181–186Google Scholar
  34. Yu H, Whittaker JW (1989) Vanadate activation of bromoperoxidase from Corallina officinalis. Biochem Biophys Res Commun 160:87–92PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Tuangporn Suthiphongchai
    • 1
    Email author
  • Patcharee Boonsiri
    • 2
  • Bhinyo Panijpan
    • 3
  1. 1.Department of Biochemistry, Faculty of ScienceMahidol UniversityBangkokThailand
  2. 2.Department of Biochemistry, Faculty of MedicineKhon Kaen UniversityKhon KaenThailand
  3. 3.Institute for Innovation and Development of Learning ProcessMahidol UniversityBangkokThailand

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