Skip to main content
SpringerLink
Log in

Isolierung und Charakterisierung der Biliproteide von Rhodella violacea (Bangiophycidae)

Isolation and characterization of the biliproteins of Rhodella violacea (Bangiophycidae)

  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

The phycobiliproteins of the unicellular marine red alga, Rhodella violacea, were investigated. A method of mass culture of the alga is presented. B-phycoerythrin, C-phycocyanin, and allophycocyanin were isolated from crude extracts by the combination of gel and ion exchange chromatography. The highly purified native phycobiliproteins were characterized by absorption spectra, absorption ratios, and fluorescence emission spectra, molecular weights, and isoelectric points. The latter were found to be 4.39, 4.46, and 4.79 for B-phycoerythrin I, C-phycocyanin, and allophycocyanin, respectively. The phycobiliprotein composition clearly distinguishes Rhodella violacea from Porphyridium aerugineum and P. cruentum, thus, supporting the separation of Rhodella violacea from the genus Porphyridium recently proposed.

Zusammenfassung

  1. 1.

    Die Zusammensetzung des Phycobiliproteidgehalts der einzelligen marinen Rotalge Rhodella violacea wurde untersucht.

  2. 2.

    Eine Methode zur Massenanzucht der Alge wird vorgestellt.

  3. 3.

    Die Isolierung und Reinigung der Phycobiliproteide aus Rohextrakten im Anschluß an die Ultraschallfraktionierung der Zellen erfolgte durch Gel- und Ionenaustauschchromatographie.

  4. 4.

    Das Vorkommen von B-Phycoerythrin, C-Phycocyanin und Allophycocyanin konnte für Rhodella violacea nachgewiesen werden.

  5. 5.

    Der hohe Reinheitsgrad der isolierten nativen Phycobiliproteide erlaubte ihre Charakterisierung durch Spektraldaten (Absorptionsspektren, Extinktionsverhältnisse und Fluorescenz-Emissionsspektren) und durch Bestimmung ihrer Molekulargewichte und isoelektrischen Punkte; die letzteren wurden mit 4,39, 4,46 und 4,79 für B-Phycoerythrin I, C-Phycocyanin bzw. Allophycocyanin ermittelt.

  6. 6.

    Die Phycobiliproteidkomposition unterscheidet Rhodella violacea deutlich von den Arten der Gattung Porphyridium und unterstützt damit die kürzlich vorgeschlagene Trennung von dieser Gattung.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

PE:

Phycoerythrin

PC:

Phycocyanin

APC:

Allophycocyanin

MG:

Molekulargewicht

IEP:

Isoelektrischer Punkt

E:

Extinktion

Literatur

  • Andrews, P.: Estimation of the molecular weights of proteins by Sephadex gelfiltration. Biochem. J. 91, 222–223 (1964)

    Google Scholar 

  • Andrews, P.: The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem. J. 96, 595–606 (1965)

    Google Scholar 

  • Bennett, A., Bogorad, L.: Properties of subunits and aggregates of blue-green algal biliproteins. Biochemistry 10, 3625–3634 (1971)

    Google Scholar 

  • Bergeron, J. A.: Studies of the localization, physicochemical properties, and action of phycocyanin in Anacystis nidulans. In: Photosynthetic mechanisms of green plants, Publ. 1145, Natl. Acad. Sci.-Natl. Research Council, pp. 527–536 (1963)

  • Berns, D. S.: Protein aggregation in phycocyanin-osmotic pressure studies. Biochem. biophys. Res. Commun. 38, 65–73 (1970)

    Google Scholar 

  • Berns, D. S., Scott, E., O'Reilly, K. T.: C-phycocyanin: minimum molecular weight. Science 145, 1054–1056 (1964)

    Google Scholar 

  • Binder, A., Wilson, K., Zuber, H.: C-phycocyanin from the thermophilic blue-green alga Mastigocladus laminosus. Isolation, characterization, and subunit composition. FEBS Letters 20, 111–116 (1972)

    Google Scholar 

  • Bloomfield, V. A., Jennings, B. R.: Molecular weight and shape of phycocyanin monomer and aggregates. Biopolymers 8, 297–299 (1969)

    Google Scholar 

  • Butler, W. L., Siegelman, H. W., Miller, C. O.: Denaturation of phytochrome. Biochemistry 3, 851–857 (1964)

    Google Scholar 

  • Chapman, D. J.: Biliproteins and bile pigments. In: N. G. Carr, B. A. Whitton, eds., The biology of blue-green algae, pp. 162–185. Oxford-London: Blackwell 1973

    Google Scholar 

  • Chapman, D. J., Cole, W. J., Siegelman, H. W.: Chromophores of allophycocyanin and R-phycocyanin. Biochem. J. 105, 903–905 (1967)

    Google Scholar 

  • Evans, L. V.: Electron microscopical observations on a new red algal unicell, Rhodella maculata, gen. nov., sp. nov. Brit. phycol. J. 5, 1–13 (1970)

    Google Scholar 

  • Förster, T.: Fluoreszenz organischer Verbindungen. Göttingen: Vandenhoeck u. Ruprecht 1951

    Google Scholar 

  • French, C. S., Smith, J. H. C., Virgin, H. I., Airth, R. L.: Fluorescencespectrum curves of chlorophylls, phaeophytins, phycoerythrins, phycocyanins, and hypericin. Plant Physiol. 31, 369–374 (1956)

    Google Scholar 

  • French, C. S., Young, V. K.: The fluorescence spectra of red algae and the transfer of energy from phycoerythrin to phycocyanin and chlorophyll. J. gen. Physiol. 35, 873–890 (1952)

    Google Scholar 

  • Fujimori, E., Pecci, J.: Distinct subunits of phycoerythrin from Porphyridium cruentum and their spectral characteristics. Arch. Biochem. Biophys. 118, 448–455 (1967)

    Google Scholar 

  • Fujimori, E., Pecci, J.: pH-dependent association of the phycoerythrin subunit. Biochim. biophys. Acta (Amst.) 207, 259–261 (1970)

    Google Scholar 

  • Fujita, Y., tsuji, T.: Photochemically active chromoprotein isolated from the blue-green alga Anabaena cylindrica. Nature (Lond.) 219, 1270–1271 (1968)

    Google Scholar 

  • Fujiwara, T.: Sugar components of phycoerythrin in Porphyra tenera. In: Ad. Davy de Virville, J. Feldmann, eds., Proc. IVth Int. Seawead Symposium, pp. 312–318. Oxford: Pergamon Press 1964

    Google Scholar 

  • Gantt, E.: Properties and ultrastructure of phycoerythrin from Porphyridium cruentum. Plant Physiol. 44, 1629–1638 (1969)

    Google Scholar 

  • Gantt, E., Conti, S. F.: Granules associated with the chloroplast lamellae of Porphyridium cruentum. J. Cell Biol. 29, 423–434 (1966)

    Google Scholar 

  • Gantt, E., Conti, S. E.: Phycobiliprotein localization in algae. Brookhaven Symp. Biol. 19, 393–405 (1967)

    Google Scholar 

  • Gantt, E., Conti, S., Conti, E.: Ultrastructure of blue-green algae. J. Bact. 97, 1486–1493 (1969)

    Google Scholar 

  • Gantt, E., Lipschultz, C. A.: Energy transfer in phycobilisomes from phycoerythrin to allophycocyanin. Biochim. biophys. Acta (Amst.) 292, 858–861 (1973)

    Google Scholar 

  • Glazer, A. N., Cohen-Bazire, G.: Subunit structure of the phycobiliproteins of blue-green algae. Proc. nat. Acad. Sci. (Wash.) 68, 1398–1401 (1971)

    Google Scholar 

  • Glazer, A. N., Cohen-Bazire, G., Stanier, R. Y.: Characterization of phycoerythrin from a Cryptomonas sp. Arch. Mikrobiol. 80, 1–18 (1971)

    Google Scholar 

  • Glazer, A. N., Fang, S.: Formation of hybrid proteins from the α and β subunits of phycocyanins of unicellular and filamentous blue-green algae. J. biol. Chem. 248, 663–671 (1973)

    Google Scholar 

  • Grabowski, J.: Chlorophyllin and phycocyanin spectra at room and low temperature. Photosynthetica 6, 291–297 (1972)

    Google Scholar 

  • Habig, W., Berns, D. S.: Isoelectric focusing of C-phycocyanins. Fed. Proc. 31, 863 Abs. (1973)

    Google Scholar 

  • Haglund, H.: Isoelectric focusing in pH-gradients. — A technique for fractionation and characterization of ampholytes. In: D. Glick, ed., Methods of biochemical analysis, Vol. 19, pp. 1–104. New York-London-Sidney: Wiley 1970

    Google Scholar 

  • Hattori, A., Crespi, H. L., Katz, J. J.: Association and dissociation of phycocyanin and the effects of deuterium substitution on the processes. Biochemistry 4, 1225–1238 (1965)

    Google Scholar 

  • Hattori, A., Fujita, Y.: Crystalline phycobilin chromoproteins obtained from a blue-green alga, Tolypothrix tenuis. J. Biochem. (Tokyo) 46, 633–644 (1959)

    Google Scholar 

  • Haxo, F. T., O'hEocha, C.: Chromoproteins of algae. In: W. Ruhland, ed., Handbuch d. Pflanzenphysiologie, Vol. 6, pp. 497–510. Berlin-Göttingen-Heidelberg: Springer 1960

    Google Scholar 

  • Jones, R. F., Speer, H. L., Kury, W.: Studies on the growth of the red alga Porphyridium cruentum. Physiol. Plant. (Copenh.) 16, 636–643 (1963)

    Google Scholar 

  • Kao, O. H., Berns, D. S., Town, W. R.: The characterization of C-phycocyanin from an extremely halo-tolerant blue-green alga, Coccochloris elabens. Biochem. J. 131, 39–50 (1973)

    Google Scholar 

  • Kobayashi, Y., Siegelman, H. W., Hirs, C. H. W.: C-phycocyanin from Phormidium luridum. Isolation of subunits. Arch. Biochem. Biophys. 152, 187–98 (1972)

    Google Scholar 

  • Kornmann, P.: Porphyridium violaceum, eine marine neue Art. Helgol. Wiss. Meeresunters. 12, 420–423 (1965)

    Google Scholar 

  • Kuhl, A., Lorenzen, H.: Handling and culturing of Chlorella. In: D. M. Prescott, ed., Methods of cell physiol., Vol. 1, pp. 159–187. New York-London: Academic Press 1964

    Google Scholar 

  • Leibo, S. P., Jones, R. F.: Freezing of the chromoprotein phycoerythrin from the red alga Porphyridium cruentum. Arch. Biochem. Biophys. 106, 78–88 (1964)

    Google Scholar 

  • Lichtlé, Chr., Giraud, G.: Aspects ultrastructuraux particuliers au plaste du Batrachospermum virgatum (Sirdt) — Rhodophycée — Nemalionale. J. Phycol. 6, 281–289 (1970)

    Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, L. A., Randall, R. J.: Protein measurement with the folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • MacColl, R., Berns, D. S., Koven, N. L.: Effect of salts on C-phycocyanin. Arch. Biochem. Biophys. 146, 477–482 (1971a)

    Google Scholar 

  • MacColl, R., Lee, J. J., Berns, D. S.: Protein aggregation in C-phycocyanin. Studies at very low concentrations with the photoelectric scanner of the ultracentrifuge. Biochem. J. 122, 421–426 (1971b)

    Google Scholar 

  • Neufeld, G. J., Riggs, A. F.: Aggregation properties of C-phycocyanin from Anacystis nidulans. Biochim. biophys. Acta (Amst.) 181, 234–243 (1969)

    Google Scholar 

  • Nolan, D. N., O'hEocha, C.: Determination of molecular weights of algal biliproteins by gel-filtration. Biochem. J. 103, 39–30 (1967)

    Google Scholar 

  • O'Carra, P., O'hEocha, C.: R-phycocyanin, a distinct type of biliprotein. Phytochemistry 4, 635–638 (1965)

    Google Scholar 

  • O'hEocha, C.: Phycobilins. In: R. A. Lewin, ed., Physiology and biochemistry of algae, pp. 421–435. New York: Academic Press 1962

    Google Scholar 

  • O'hEocha, C.: Biliproteins. In: T. W. Goodwin, ed., Biochemistry of chloroplasts, Vol. 1, pp. 407–421. New York-London: Academic Press 1966

    Google Scholar 

  • Rüdiger, W.: Gallenfarbstoffe und Biliproteide. In: W. Herz, H. Grisebach, G. W. Kirby, eds., Fortschr. d. Chemie organ. Naturstoffe, Vol. 29, pp. 60–139. Wien-New York: Springer 1971

    Google Scholar 

  • Schram, B. L., Kroes, H. H.: Structure of phycocyanobilin. Europ. J. Biochem. 19, 581–594 (1971)

    Google Scholar 

  • Teale, F. W. J., Dale, R. E.: Isolation and spectral characterization of phycobiliproteins. Biochem. J. 116, 161–169 (1970)

    Google Scholar 

  • Tiselius, A., Hjertén, Levin, Ö: Protein chromatography on calcium phosphate columns. Arch. Biochem. Biophys. 65, 132–155 (1956)

    Google Scholar 

  • Torjesen, P. A., Sletten, K.: C-phycocyanin from Oscillatoria agardhii. I. Some molecular properties. Biochim. biophys. Acta (Amst.) 263, 258–271 (1972)

    Google Scholar 

  • Urbach, W., Rupp, W., Sturm, H.: Experimente zur Stoffwechselphysiologie der Pflanzen, Teil 1. Botanisches Institut I der Universität Würzburg, Selbstverlag (1972)

  • Van der Velde, H. H.: The natural occurrence in red algae of two phycoerythrins with different molecular weights and spectral properties. Biochim. biophys. Acta (Amst.) 303, 246–257 (1973)

    Google Scholar 

  • Vernotte, C.: Séparation et caractérisation spectroscopique du monomère et des polymères de la C-phycocyanine. Photochem. and Photobiol. 14, 163–173 (1971)

    Google Scholar 

  • Vesterberg, O.: Isoelectric focusing of proteins. Inaugural-Dissertation, Stockholm (1968)

  • Wehrmeyer, W.: Elektronenmikroskopische Untersuchung zur Feinstruktur von Porphyridium violaceum Kornmann mit Bemerkungen über seine taxonomische Stellung. Arch. Mikrobiol. 75, 121–139 (1971)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Fachbereich Biologie-Botanik der Universität Marburg a. d. Lahn, Marburg a. d. Lahn, Deutschland

    Klaus P. Koller & Werner Wehrmeyer

Authors
  1. Klaus P. Koller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Werner Wehrmeyer
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Koller, K.P., Wehrmeyer, W. Isolierung und Charakterisierung der Biliproteide von Rhodella violacea (Bangiophycidae). Arch. Microbiol. 100, 253–270 (1974). https://doi.org/10.1007/BF00446322

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00446322

Key words

Search

Navigation