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Purification and Characterization of Cadmium-Binding Protein from Unicelluar Alga Chlorella sorokinian

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Abstract

The unicellular green alga Chlorella sorokiniana ANA9 is highly resistant to heavy metals, and its metal-binding proteins are induced in the presence of cadmium. A novel cadmium-binding protein in C. sorokiniana cultured in 100 mg/l cadmium ions for 4 days was isolated and characterized. The crude protein extract was obtained by cell disruption and partly purified by ammonium sulfate precipitation. After purification by anion-exchange chromatography with diethylaminoethyl (DEAE)-Sepharose CL-6B, the protein was further purified by gel filtration with Sephacryl S-100, followed by Sephadex G-75. The molecular weight of the purified protein was determined to be 11.5 kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The cadmium binding capacity of the purified protein was 119 μg/mg. The involvement of thiol coordination in metal-ion binding was confirmed by measuring the ultraviolet spectrum. This article is the first to describe the metallothionein-like cadmium-binding protein from Chlorella species, the expression of which is induced by cadmium exposure.

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Literature Cited

  1. Coupe SA, Taylor JE, Roberts JA (1995) Characterisation of an mRNA encoding a metallothionein-like protein that accumulates during ethylene-promoted abscission of Sambucus nigra L. leaflets. Planta 197:442–447

    Article  PubMed  CAS  Google Scholar 

  2. de Framond AJ (1991) A metallothionein-like gene from maize (Zea mays). Cloning and characterization. FEBS Lett 290:103–106

    Article  PubMed  Google Scholar 

  3. Evans IM, Gatehouse LN, Gatehouse JA, Robinson NJ, Croy RR (1990) A gene from pea (Pisum sativum L.) with homology to metallothionein genes. FEBS Lett 262:29–32

    Article  PubMed  CAS  Google Scholar 

  4. Gouvea SP, Vieira AA, Lombardi AT (2005) Copper and cadmium complexation by high molecular weight materials of dominant microalgae and of water from a eutrophic reservoir. Chemosphere 60:1332–1339

    Article  PubMed  CAS  Google Scholar 

  5. Hamer DH (1986) Metallothionein. Annu Rev Biochem 55:913–51

    PubMed  CAS  Google Scholar 

  6. 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 

  7. Le Faucheur S, Behra R, Sigg L (2005) Phytochelatin induction, cadmium accumulation, and algal sensitivity to free cadmium ion in Scenedesmus vacuolatus. Environ Toxicol Chem 24:1731–1737

    Article  PubMed  CAS  Google Scholar 

  8. Lodeiro P, Cordero B, Barriada JL, Herrero R, Sastre de Vicente ME (2005) Biosorption of cadmium by biomass of brown marine macroalgae. Bioresour Technol 96:1796–1803

    Article  PubMed  CAS  Google Scholar 

  9. Morris CA, Nicolaus B, Sampson V, Harwood JL, Kille P (1999) Identification and characterization of a recombinant metallothionein protein from a marine alga, Fucus vesiculosus. Biochem J 338:553–560

    Article  PubMed  CAS  Google Scholar 

  10. Nietfeld W, el-Baradi T, Mentzel H, Pieler T, Koster M, Poting A, et al. (1989) Second-order repeats in Xenopus laevis finger proteins. J Mol Biol 208:639–659

    Article  PubMed  CAS  Google Scholar 

  11. Parkhill J, Sebaihia M, Preston A, Murphy LD, Thomson N, Harris DE, et al. (2003) Comparative analysis of the genome sequences of Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica. Nat Genet 35:32–40

    Article  PubMed  Google Scholar 

  12. Robinson NJ, Tommey AM, Kuske C, Jackson PJ (1993) Plant metallothioneins. Biochem J 295:1–10

    PubMed  CAS  Google Scholar 

  13. Schmidt T, Schlegel HG (1994) Combined nickel-cobalt-cadmium resistance encoded by the ncc locus of Alcaligenes xylosoxidans 31A. J Bacteriol 176:7045–7054

    PubMed  CAS  Google Scholar 

  14. Yoshida N, Ikeda R, Okuno T (2006) Identification and characterization of heavy metal-resistant unicellular alga isolated from soil and its potential for phytoremediation. Bioresour Technol (in press)

  15. Zhou J, Goldsbrough PB (1994) Functional homologs of fungal metallothionein genes from Arabidopsis. Plant Cell 6:875–884

    Article  PubMed  CAS  Google Scholar 

  16. Zhou J, Goldsbrough PB (1995) Structure, organization and expression of the metallothionein gene family in Arabidopsis. Mol Gen Genet 248:318–328

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry and Applied Biosciences, University of Miyazaki, 1-1 Gakuen Kibanadai-Nishi, Miyazaki, 889-2192, Japan

    Naoto Yoshida, Kazushige Ishii, Tomoko Okuno & Kazunori Tanaka

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  1. Naoto Yoshida
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  2. Kazushige Ishii
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  3. Tomoko Okuno
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  4. Kazunori Tanaka
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Correspondence to Naoto Yoshida.

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Yoshida, N., Ishii, K., Okuno, T. et al. Purification and Characterization of Cadmium-Binding Protein from Unicelluar Alga Chlorella sorokinian. Curr Microbiol 52, 460–463 (2006). https://doi.org/10.1007/s00284-005-0328-z

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  • DOI: https://doi.org/10.1007/s00284-005-0328-z

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