Skip to main content
SpringerLink
Log in

The genome of the thermoacidophilic red microalga Galdieria sulphuraria encodes a small family of secreted class III peroxidases that might be involved in cell wall modification

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

We report the identification of a small family of secreted class III plant peroxidases (Prx) from the genome of the unicellular thermoacidophilic red alga Galdieria sulphuraria (Cyanidiaceae). Apart from two class I ascorbate peroxidases and one cytochrome c peroxidase, the red algal genome encodes four class III plant peroxidases, thus complementing the short list of algal cell wall peroxidases (Passardi et al. in Genomics 89:567–579, 2007). We have characterized the family gene structure, analyzed the extracellular space and cell wall fraction of G. sulphuraria for the presence of peroxidase activity and used shotgun proteomics to identify candidate extracellular peroxidases. For a detailed enzymatic characterization, we have purified a secreted peroxidase (GsPrx04) from the cell-free medium using hydrophobic interaction chromatography. The enzyme proved heat and acid-stable and exhibited an apparent molecular mass of 40 kDa. Comparative genomics between endolithically growing G. sulphuraria and a close relative, the obligatory aquatic, cell wall-less Cyanidioschyzon merolae, revealed that class III peroxidases only occur in the terrestrial microalga, thus supporting the key function of these enzymes in the process of land colonization.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

Prx:

Peroxidase

References

  • Bailey RW, Staehelin LA (1968) The chemical composition of isolated cell walls of Cyanidium caldarium. J Gen Microbiol 54:269–276

    PubMed  CAS  Google Scholar 

  • Bakalovic N, Passardi F, Ioannidis V, Cosio C, Penel C, Falquet L, Dunand C (2006) PeroxiBase: a class III plant peroxidase database. Phytochemistry 67:534–549

    Article  PubMed  CAS  Google Scholar 

  • Barbier G, Oesterhelt C, Larson MD, Halgren RG, Wilkerson C, Gravito M, Benning C, Weber APM (2005) Comparative genomics of two closely related unicellular thermo-acidophilic red algae, Galdieria sulphuraria and Cyanidioschyzon merolae, reveals the molecular basis of the metabolic flexibility of G. sulphuraria and significant differences in carbohydrate metabolism of both algae. Plant Physiol 137:460–474

    Article  PubMed  CAS  Google Scholar 

  • Bhattacharya D, Yoon H, Hackett J (2004) Photosynthetic eukaryotes unite: endosymbiosis connects the dots. Bioessays 26:50–60

    Article  PubMed  Google Scholar 

  • Blum H, Beier H, Gross HJ (1987) Improved silver staining for plant proteins, RNA, and DNA in polyacrylamide gels. Electrophoresis 8:93–99

    Article  CAS  Google Scholar 

  • 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 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Ciniglia C, Yoon HS, Pollio A, Pinto G, Bhattacharya D (2004) Hidden biodiversity of the extremophilic Cyanidiales red algae. Mol Ecol 13:1827–1838

    Article  PubMed  CAS  Google Scholar 

  • Cosgrove DJ (2001) Wall structure and wall loosening. A look backwards and forwards. Plant Physiol 125:131–134

    Article  PubMed  CAS  Google Scholar 

  • DeLuca P, Musacchio A, Taddei R (1972) Diversio comportamento in eterotrofia delle due forme de Cyandium caldarium dei Campi Flegrei (Napoli). Delphinoa 12/13:19–27

    Google Scholar 

  • Doemel WN, Brock TD (1970) The upper temperature limit of Cyanidium caldarium. Arch Microbiol 72:326–332

    CAS  Google Scholar 

  • Duroux L, Welinder KG (2003) The peroxidase gene family in plants: a phylogenetic overview. J Mol Evol 57:397–407

    Article  PubMed  CAS  Google Scholar 

  • Enami I, Akutsu H, Kyogoku Y (1986) Intracellular pH regulation in an acidophilic unicellular alga—Cyanidium caldarium—P-31-NMR determination of intracellular pH. Plant Cell Physiol 27:1351–1359

    CAS  Google Scholar 

  • Encina A, Fry SC (2005) Oxidative coupling of a feruloyl-arabinoxylan trisaccharide (FAXX) in the walls of living maize cells requires endogenous hydrogen peroxide and is controlled by a low-Mr apoplastic inhibitor. Planta 223:77–89

    Article  PubMed  CAS  Google Scholar 

  • Fry SC (1998) Oxidative scission of plant cell wall polysaccharides by ascorbate-induced hydroxyl radicals. Biochem J 332:507–515

    PubMed  CAS  Google Scholar 

  • Gajhede M, Schuller DJ, Henriksen A, Smith AT, Poulos TL (1997) Crystal structure of horseradish peroxidase C at 2.15 A resolution. Nat Struct Biol 12:1032–1038

    Article  Google Scholar 

  • Gasteiger E, Hoogland C, Gattiker A, Duvaud D, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the ExPASy Server. In: Walker JM (ed) The proteomics protocols handbook. Humana, Totowa, pp 571–607

    Google Scholar 

  • Gray JSS, Montgomery R (2006) Asymmetric glycosylation of soybean seed coat peroxidase. Carbohydr Res 341:198–209

    Article  PubMed  CAS  Google Scholar 

  • Gross W (2000) Ecophysiology of algae living in highly acidic environments. Hydrobiologia 433:31–37

    Article  CAS  Google Scholar 

  • Gross W, Oesterhelt C (1999) Ecophysiological studies on the red alga Galdieria sulphuraria isolated from Southwest Iceland. Plant Biol 1:694–700

    Google Scholar 

  • Gross W, Schnarreberger C (1995) Heterotrophic growth of two strains of the acido-thermophilic red alga Galdieria sulphuraria. Plant Cell Physiol 36:633–638

    CAS  Google Scholar 

  • Gross W, Küver J, Tischendorf G, Bouchaala N, Büsch W (1998) Cryptoendolithic growth of the red alga Galdieria sulphuraria in volcanic areas. Eur J Phycol 33:25–31

    Article  Google Scholar 

  • Haebel S, Kehr J (2001) Matrix-assisted laser desorption/ionization time of flight mass spectrometry peptide mass fingerprints and post source decay: a tool for the identification and analysis of phloem proteins from Cucurbita maxima Duch. separated by two-dimensional polyacrylamide gel electrophoresis. Planta 213:586–593

    Article  PubMed  CAS  Google Scholar 

  • Henriksen A, Mirza O, Indiani C, Teilum K, Smulevich G, Welinder KG, Gajhede M (2001) Structure of soybean seed coat peroxidase: a plant peroxidase with unusual stability and haem–apoprotein interactions. Prot Sci 10:108–115

    Article  CAS  Google Scholar 

  • Hiraga S, Sasaki K, Ito H, Ohashi Y, Mtsui H (2001) A large family of class III plant peroxidases. Plant Cell Physiol 42:462–468

    Article  PubMed  CAS  Google Scholar 

  • Jansson S, Stefansson H, Nystrom U, Gustafsson P, Albertsson PA (1997) Antenna protein composition of PS I and PS II in thylakoid sub-domains. Biochim Biophys Acta 1320:297–309

    Article  CAS  Google Scholar 

  • Kawano T (2003) Roles of the reactive oxygen species-generating peroxidase reactions in plant defense and growth induction. Plant Cell Rep 21:829–837

    PubMed  CAS  Google Scholar 

  • Keller A, Nesvizhskii AI, Kolker E, Aebersold R (2002) Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem 74:5383–5392

    Article  PubMed  CAS  Google Scholar 

  • Kerr EM, Fry SC (2004) Extracellular cross-linking of xylan and xyloglucan in maize cell-suspension cultures: the role of oxidative phenolic coupling. Planta 219:73–83

    Article  PubMed  CAS  Google Scholar 

  • Kitajima S, Ueda M, Sano S, Mixake C, Hohchi T, Tomizawa K, Shigeoka S, Yokota A (2002) Stable form of ascorbate peroxidase from the red alga Galdieria partita similar to both chloroplastic and cytosolic isoforms of higher plants. Biosci Biotechnol Biochem 66:2367–2375

    Article  PubMed  CAS  Google Scholar 

  • Kitajima S, Tomizawa K, Shigeoka S, Yokota A (2006) An inserted loop region of stromal ascorbate peroxidase is involved in its hydrogen peroxide-mediated inactivation. FEBS J 273:2704–2710

    Article  PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Li ZC, McClure W, Hagerman AE (1989) Soluble and bound apoplastic activity for peroxidase, beta-d-glucosidase, malate dehydrogenase and nonspecific arylesterase in barley (Hordeum vulgare L.) and oat (Avena sativa L.) primary leaves. Plant Physiol 90:185–190

    Article  PubMed  CAS  Google Scholar 

  • Matsuzaki M, Misumi O, Shin-I T, Maruyama S, Takahara M, Miyagishima SY, Mori T, Nishida K, Yagisawa F, Nishida K, Yoshida Y, Nishimura Y, Nakao S, Kobayashi T, Momoyama Y, Higashiyama T, Minoda A, Sano M, Nomoto H, Oishi K, Hayashi H, Ohta F, Nishizaka S, Haga S, Miura S, Morishita T, Kabeya Y, Terasawa K, Suzuki Y, Ishii Y, Asakawa S, Takano H, Ohta N, Kuroiwa H, Tanaka K, Shimizu N, Sugano S, Sato N, Nozaki H, Ogasawara N, Kohara Y, Kuroiwa T (2004) Genome sequence of the ultrasmall unicellular red alga Cyanidioschyzon merolae 10D. Nature 428:653–657

    Article  PubMed  CAS  Google Scholar 

  • McEldoon JP, Pokora AR, Dordick JS (1995) Lignin peroxidase-type activity of soybean peroxidase. Enzyme Microb Technol 17:359–365

    Article  CAS  Google Scholar 

  • McEldoon JP, Dordick JS (1996) Unusual thermal stability of soybean peroxidase. Biotechnol Prog 12:555–558

    Article  CAS  Google Scholar 

  • Merola A, Castaldo R, DeLuca P, Gambardella R, Musacchio A, Taddei R (1981) Revision of Cyanidium caldarium. Three species of acidophilic algae. G Bot Ital 115:189–195

    Google Scholar 

  • Nesvizhskii AI, Keller A, Kolker E, Aebersold R (2003) A statistical model for identifying proteins by tadem mass spectrometry. Anal Chem 75:4646–4658

    Article  PubMed  CAS  Google Scholar 

  • Nissum M, Schiødt CB, Welinder KG (2001) Reactions of soybean peroxidase and hydrogen peroxide pH 2,4–12,0, and veratryl alcohol at pH 2,4. Biochim Biophys Acta 1545:339–348

    PubMed  CAS  Google Scholar 

  • Oesterhelt C, Schnarreberger C, Gross W (1999) Characterization of a sugar/polyol-uptake system in the red alga Galdieria sulphuraria. Eur J Phycol 34:271–277

    Google Scholar 

  • Passardi F, Longet D, Penel C, Dunand C (2004a) The plant peroxidase multigenic family in rice and its evolution in green plants. Phytochemistry 65:1879–1893

    Article  PubMed  CAS  Google Scholar 

  • Passardi F, Penel C, Dunand C (2004b) Performing the paradoxical: how plant peroxidases modify the cell wall. Trends Plant Sci 9:534–540

    Article  PubMed  CAS  Google Scholar 

  • Passardi F, Cosio C, Penel C, Dunand C (2005) Peroxidases have more functions than a Swiss army knife. Plant Cell Rep 24:255–265

    Article  PubMed  CAS  Google Scholar 

  • Passardi F, Bakalovic N, Teixeira FK, Margis-Pinheiro M, Penel C, Dunand C (2007) Prokaryotic origins of the non-animal peroxidase superfamily and organelle-mediated transmission to eukaryotes. Genomics 89:567–579

    Article  PubMed  CAS  Google Scholar 

  • Sano S, Ueda M, Kitajima S, Takeda T, Shigeoka S, Kurano N, Miyachi S, Miyake C, Yokota A (2001) Characterization of ascorbate peroxidase from unicellular red alga Galdieria partita. Plant Cell Physiol 42:433–440

    Article  PubMed  CAS  Google Scholar 

  • Tamas L, Durcekova K, Haluskova L, Huttova J, Mistrik I, Olle M (2007) Rhizosphere localized cationic peroxidase from barley roots is strongly activated by cadmium and correlated with root growth inhibition. Chemosphere 66:1292–1300

    Article  PubMed  CAS  Google Scholar 

  • Trivedi UJ, Bassi AS, Zhu JX (2006) Investigation of phenol removal using sol-gel/alginate immobilized soybean seed hull peroxidase. Can J Chem Eng 84:239–247

    Article  CAS  Google Scholar 

  • Weber APM, Oesterhelt C, Gross W, Bräutigam A, Imboden LA, Krassovskaya I, Linka N, Truchina J, Schneidereit J, Voll LM, Zimmermann M, Riekhof WR, Yu B, Garavito RM, Benning C (2004) EST-analysis of the thermo-acidophilic red microalga Galdieria sulphuraria. Plant Mol Biol 55:17–32

    Article  PubMed  CAS  Google Scholar 

  • Weber APM, Weber KL, Carr K, Wikerson C, Ohlrogge JB (2007) Sampling the Arabidopsis transcriptome with massively-parallel pyrosequencing. Plant Physiol 144:32–42

    Article  PubMed  CAS  Google Scholar 

  • Welinder KG (1992a) Plant peroxidases: structure–function relationships. In: Penel C, Gaspar T, Greppin H (eds) Plant peroxidases. University of Geneva, Geneva, Switzerland, pp 1–24

    Google Scholar 

  • Welinder KG (1992b) Superfamily of plant, fungal and bacterial peroxidases. Curr Opin Struct Biol 2:388–393

    Article  CAS  Google Scholar 

  • Welinder KG, Mauro JM, Nørskov Lauritsen L (1992) Structure of plant and fungal peroxidases. Biochem Soc Trans 20:337–340

    PubMed  CAS  Google Scholar 

  • Welinder KG, Justesen AF, Kjærsgård IVH, Jensen RB, Rasmussen SK, Jespersen HM, Duroux L (2002) Structural diversity and transcription of class III peroxidases from Arabidopsis thaliana. Eur J Biochem 269:6063–6081

    Article  PubMed  CAS  Google Scholar 

  • Yoon H, Hackett J, Pinto G, Bhattacharya D (2002) The single, ancient origin of chromist plastids. Proc Natl Acad Sci USA 99:15507–15512

    Article  PubMed  CAS  Google Scholar 

  • Yoon H, Hackett J, Ciniglia C, Pinto G, Bhattacharya D (2004) A molecular timeline for the origin of photosynthetic eukaryotes. Mol Biol Evol 21:809–818

    Article  PubMed  CAS  Google Scholar 

  • Yoon HS, Ciniglia C, Wu M, Comeron JM, Pinto G, Pollio A, Bhattacharya D (2006) Establishment of endolithic populations of extremophilic Cyanidiales (Rhodophyta). BMC Evol Biol 6:78–90

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors would like to acknowledge financial aid from the Deutsche Forschungsgemeinschaft (Emmy Noether Grant) to C.O., by a postdoctoral fellowship of the German Academic Exchange Service (DAAD) to M.S. and a grant of the National Science Foundation (EF-0332882) to A.P.M.W. We are grateful to the Michigan State University Research Technology Support Facility for mass spectrometry-based protein identification. We would like to dedicate this paper to the memory of the late Dr. Wolfgang Gross who initiated the work on extracellular proteins of Galdieria sulphuraria.

Author information

Author notes

  1. S. Vogelbein

    Present address: Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Str. 10, 13125, Berlin, Germany

  2. A. P. M. Weber

    Present address: Institut für Biochemie der Pflanzen, Heinrich-Heine-Universität, Universitätsstr. 1, 40225, Düsseldorf, Germany

Authors and Affiliations

  1. Institut für Biochemie und Biologie, Universität Potsdam, Karl-Liebknecht-Str. 24-25, Haus 20, 14476, Potsdam-Golm, Germany

    C. Oesterhelt

  2. Institut für Biologie, Freie Universität Berlin, Königin-Luise-Str. 12-16, 14197, Berlin, Germany

    S. Vogelbein

  3. Department of Plant Biology, Michigan State University, East Lansing, MI, 48824, USA

    R. P. Shrestha & A. P. M. Weber

  4. Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Abteilung Bioinformatik, Goldschmidtstr. 1, 37077, Göttingen, Germany

    M. Stanke

Authors
  1. C. Oesterhelt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Vogelbein
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. P. Shrestha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Stanke
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. P. M. Weber
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to C. Oesterhelt.

Additional information

Nucleotide sequence database accession numbers: GsuAPX01 (EF589723), GsuAPX02 (EF589721), GsuCcP01 (EF589722), GsPrx01 (EF589724), GsPrx02 (EF589725), GsPrx03 (EF589726), and GsPrx04 (EF589727). The nomenclature of peroxidases has been adapted to PeroxiBase (http://peroxidase.isb-sib.ch/).

Electronic supplementary material

Below is the link to the electronic supplementary material.

Fig. S1

Neighbour-joining tree of class I and class III peroxidases. The phylogenetic tree was constructed from derived protein sequences using the ClustalW and Phylip services at the Institut Pasteur (http://www.pasteur.fr/english.html). Bootstrap values are indicated on the branches. Galdieria sulphuraria sequences, generated in this study, are indicated by an asterisk. Proteins from organisms other than G. sulphuraria were taken from the PeroxiBase (http://peroxidase.isb-sib.ch/; Bakolovic et al. 2006). For those proteins available at NCBI (http://www.ncbi.nlm.nih.gov/), the corresponding accession numbers are also given in parentheses. AruPrx01a (AAA33377) corresponds to horseradish peroxidase HRP C1, generally used as reference for class III peroxidases. The putative cytochrome c peroxidase from G. sulphuraria (GsuCcP01) was used as an outgroup (PDF 17 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Oesterhelt, C., Vogelbein, S., Shrestha, R.P. et al. The genome of the thermoacidophilic red microalga Galdieria sulphuraria encodes a small family of secreted class III peroxidases that might be involved in cell wall modification. Planta 227, 353–362 (2008). https://doi.org/10.1007/s00425-007-0622-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-007-0622-z

Keywords

Search

Navigation