Skip to main content
SpringerLink
Log in

Purification and characterization of a novel haemagglutinin from Chlorella pyrenoidosa

  • Original Paper
  • Published:
Journal of Industrial Microbiology and Biotechnology

Abstract

We previously identified a strong haemagglutination activity in the freshwater unicellular green alga, Chlorella pyrenoidosa. Here, we sought to purify and characterize the haemagglutinin associated with this activity. Ammonium sulfate precipitation, gel filtration on sephacryl S-200 and DEAE-Sepharose ion-exchange chromatography were used to purify the haemagglutinin, which was designated CPH (Chlorella pyrenoidosa haemagglutinin). The molecular weight of CPH was estimated as 58 kDa by SDS-PAGE and 60 kDa by gel filtration of the native protein, indicating that this haemagglutinin exists as a monomer. The haemagglutinin activity of CPH was inhibited by glycoproteins, especially yeast mannan, but not by monosaccharides or disaccharides, indicating that CPH is carbohydrate-specific. In addition to the composition of CPH shown to be rich in glycine and acidic amino acids, heamagglutinating activity of CPH was insensitive to variations in pH or the presence of divalent cations, and atomic force microscopy revealed that the protein is rod-shaped. These results indicate that the characteristics of CPH are consistent with its identification as a haemagglutinin, and suggest that CPH may be a viable candidate for applications in a variety of biomedical fields.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Ainouz IL, Simpaio H, Benevides NMB, Freitas ALP, Costa FHF, Carvalho MR. Joventino FP (1992) Agglutination of enzyme treated erythrocytes by Brazilian marine algal extracts. Bot Mar 35:475–479

    CAS  Google Scholar 

  2. Benevides NM, Holanda ML, Melo FR, Freitas ALP, Simpaio H (1998) Purification and partial characterization of the lectin from marine red alga Enantiocladia duperreyi (C. Agardh) Falkenberg. Bot Mar 41:521–525

    Article  CAS  Google Scholar 

  3. Boyd WC, Almodovar LR, Boyd JG (1966) Agglutinins from marine algae for human erythrocytes. Transfusion (Philadelphia) 6:82–83

    CAS  Google Scholar 

  4. Chu CY, Laio WR, Huang R, Lin LP (2004) Hemagglutinating and antibiotic activities of freshwater microalgae. World J Microbiol Biotechnol 20:817–825

    Article  CAS  Google Scholar 

  5. Chiles TC, Bird KT (1990) Gracilaria tikvahiae agglutinin: partial purification and purification and preliminary characterization of its carbohydrate specificity. Carbohydr Res 207:319–326

    Article  CAS  Google Scholar 

  6. Dudois M, Gilles KA, Hamilton JK, Rogers PA, Smith F (1956) Colorimetric method for determination of sugar and related substances. Anal Chem 28:350

    Article  Google Scholar 

  7. Fabregas J, Llovo J, Munoz A (1985) Hemagglutinins in red seaweeds. Bot Mar 28:517–520

    CAS  Google Scholar 

  8. Freitas ALP, Teixeira DIA, Costa FHF, Farias WRL, Lobato ASC, Sampaio AH, Benevides NMB (1997) A new survey of Brazilian marine algae for agglutinins. J Appl Phycol 9:495–501

    CAS  Google Scholar 

  9. Hori K, Miyazawa K, lto K (1986) Preliminary characterization of agglutinins from seven marine algal species. Bull Jpn Soc Fish 52:323–331

    Google Scholar 

  10. Hoir K, Oiwa C, Miyazawa K, Ito K (1988) Evidence for wide distribution of agglutinins. Bot Mar 31:133–138

    Article  Google Scholar 

  11. Hori K, Miyazawa K, Ito K (1990) Some common properties of lectins from marine algae. Hydrobiol 204/205:561–566

    Article  Google Scholar 

  12. Hori K, Ogata T, Kamiya H, Mimuro M (1996) Lectin-like and lectin receptors in marine microalgae hemagglutination and reactivity with purified lectins. J Phycol 32:783–790

    Article  CAS  Google Scholar 

  13. Hu CY, Lin. LP (2003) Purification and characterization of hydrolytic enzymes in Sinorhizobium fredii CCRC15769. World J Microbiol Biotechnol 19:515–522

    Article  CAS  Google Scholar 

  14. Illman AM, Scragg AH, Shales SW (2000) Increase in Chlorella stains calorific values when grown in low nitrogen medium. Enz Microbiol Tech 27:631–635

    Article  CAS  Google Scholar 

  15. Kapila WM, Nabil AA, Jayne CG, Xu S, Liu GY (2001) Fabrication of nanometer-sized protein patterns using atomic force microscopy and selective immobilization. Biophys J 80:1891–1899

    Article  Google Scholar 

  16. Kawakubo A, Makino H, Ohnishi JI, Hirohara H, Hori K (1997) The marine red algae Eucheuma serra (J. Agardh), a high yielding source of two isolectins. J Appl Phycol 9:331–338

    Article  CAS  Google Scholar 

  17. Konishi F, Mitsuyama M, Okuda M, Tanaka K, Hasegawa T, Nomoto K (1996) Protective effect of an acidic glycoprotein obtained from culture of Chlorella vulgaris against myelosuppression by 5-fluorouracil. Cancer Immunol Immunotherapy 42:268–274

    Article  CAS  Google Scholar 

  18. Liao WR, Huang R (2000) Agglutination of human and animal erythrocytes in marine unicellular algae. J Ind Microbiol Biotechnol 24:262–266

    Article  CAS  Google Scholar 

  19. Lima HC, Costa FHF, Simpaio AH, Neves SA, Benevides NMB, Teixeira DIA, Rogers DJ, Freitas ALP (1998) Induction and inhibition of human lymphocyte transformation by the lectin from the red marine algae Amansia multifida. J Appl Phycol 10:153–162

    Article  CAS  Google Scholar 

  20. Lowry OM, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265

    CAS  Google Scholar 

  21. Marchin KL, Berrie CL (2003) Conformational changes in the plasma protein fibrinogen upon adsorption to graphite and mica investigated by atomic force microscopy. Langmuir 19:9883–9888

    Article  CAS  Google Scholar 

  22. Neves SA, Baruffi MD, Freitas ALP, Barreira MCR (2001) Neutrophil migration induced in vivo and vitro by marine algae lectins. Inflam Res 50:486–490

    Article  CAS  Google Scholar 

  23. Noda K, Tanaka K, Yamada A, Ogata J, Tanaka H, Shoyama Y (2002) Simple assay for antitumour immunoactive glycoprotein derived from Chlorella vulgaris strain CK22 using ELISA. Phytother Res 16:581–585

    Article  CAS  Google Scholar 

  24. Rogers DJ, Blunden G, Topliss JA, Guiry MD (1980) A survey of some marine organisms for hemagglutinins. Bot Mar 23:569–577

    Google Scholar 

  25. Rogers DJ, Hori K (1993) Marine algal lectins: new developments. Hydrobiol 260/261:589–593

    Article  Google Scholar 

  26. Sampaio AH, Rogers DJ, Barwell CJ, Sampaio SS, Costa FHF, Ramos MV (1999) A new isolation procedure and further characterization of the lectin from the red alga Ptilota serrata. J Appl Phycol 10:539–546

    Article  Google Scholar 

  27. Sato Y, Murakami M, Miyazawa K, Hori K (2000) Purification and characterization of a novel lectin from a freshwater cyanobacterium, Oscillatoria agardhii. Comp Biochem Phys Part B 125:169–177

    Article  CAS  Google Scholar 

  28. Tanaka K, Yamada A, Noda K, Hasegawa T, Okuda M, Shoyama Y, Nomoto K (1998) A novel glycoprotein obtained from Chlorella vulgaris strain CK22 shows antimetastatic immunopotentiation. Cancer Immunol Immunother 45:313–320

    Article  CAS  Google Scholar 

  29. Yamaguchi M, Mituru J, Ryuichi S, Koji M, Kamiya H (1998) Purification and characterization of Microcystis aeruginosa (freshwater cyanobacterium) lectin. Comp Biochem Phys Part B. 119:593–597

    Article  CAS  Google Scholar 

  30. Yamaguchi M, Ogawa T, Muramoto K, Kamio Y, Jimbo M Kamiya H (1999) Isolation and characterization of mannan-binding lectin from the freshwater cyanobacterium (blue-green algae) Microcystis viridis. Biochem Biophys Res Comm 265:703–708

    Article  CAS  Google Scholar 

  31. Wu HL, Hseu RS, Lin LP (2000) Utilization of ribosomal DNA sequences to identify Chlorella spp. isolates. Bot Bull Acad Sin 42:115–121

    Google Scholar 

Download references

Acknowledgments

We wish to acknowledge the financial support for this study received from the National Science Council (Taiwan) (NSC: 92–2317-B-002–030).

Author information

Authors and Affiliations

  1. Institute of Microbiology and Biochemistry, National Taiwan University, Taipei, 106, Taiwan

    Chun-Yao Chu & Liang-Ping Ling

  2. Institute of Oceanography, National Taiwan University, Taipei, 106, Taiwan

    Chun-Yao Chu & Rang Huang

Authors
  1. Chun-Yao Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rang Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Liang-Ping Ling
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Liang-Ping Ling.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chu, CY., Huang, R. & Ling, LP. Purification and characterization of a novel haemagglutinin from Chlorella pyrenoidosa . J Ind Microbiol Biotechnol 33, 967–973 (2006). https://doi.org/10.1007/s10295-006-0145-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-006-0145-9

Keywords

Search

Navigation