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A novel rapid and continuous procedure for large-scale purification of magnetosomes from Magnetospirillum gryphiswaldense

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Abstract

A new rapid and continuous procedure was developed for purifying magnetosomes from Magnetospirillum gryphiswaldense MSR-1 cells on a large scale. The procedure included these steps: disruption of cells with a high-pressure homogeniser, isolation of magnetosomes with a continuous magnetism isolation system accompanied by low-power ultrasonication and urea treatment, removal of adsorbed and surface proteins with proteinase K, removal of nucleic acids with electro-elution, and replacement of the PBS buffer with distilled water by a magnetically stirred system. The purified magnetosomes were stored at −20 °C after lyophilized and treated with γ-rays. The time required for purification was reduced from 20–30 to 2–5 days. Evaluation of the purity of the resulting magnetosomes was carried out with SDS-PAGE, PCR, and Fourier-transform infrared spectroscopy. The overall data suggest that the method presented here is a simple, rapid, continuous, and highly efficient procedure for large-scale purification of magnetosomes.

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References

  • Arakaki A, Webb J, Matsunaga T (2003) A novel protein tightly bound to bacterial magnetic particles in Magnetospirillum magneticum strain AMB-1. J Biol Chem 278(10):8745–8750

    Article  CAS  Google Scholar 

  • Bazylinski DA, Frankel RB (2003) Magnetosome formation in prokaryotes. Nat Rev Micro 2(3):217–230

    Article  Google Scholar 

  • Blakemore R (1975) Magnetotactic bacteria. Science 190(4212):377–379

    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(1–2):248–254

    Article  CAS  Google Scholar 

  • Ceyhan B, Alhorn P, Lang C, Schüler D, Niemeyer C (2006) Semisynthetic biogenic magnetosome nanoparticles for the detection of proteins and nucleic acids. Small 2(11):1251–1255

    Article  CAS  Google Scholar 

  • Gorby YA, Beveridge TJ, Blakemore RP (1988) Characterization of the bacterial magnetosome membrane. J Bacteriol 170(2):834–841

    Article  CAS  Google Scholar 

  • Grünberg K, Wawer C, Tebo BM, Schüler D (2001) A large gene cluster encoding several magnetosome proteins is conserved in different species of magnetotactic bacteria. Appl Environ Microbiol 67(10):4573–4582

    Article  Google Scholar 

  • Grünberg K, Müller EC, Otto A, Reszka R, Linder D, Kube M, Reinhardt R, Schüler D (2004) Biochemical and proteomic analysis of the magnetosome membrane in Magnetospirillum gryphiswaldense. Appl Environ Microbiol 70(2):1040–1050

    Article  Google Scholar 

  • Heyen U, Schüler D (2003) Growth and magnetosome formation by microaerophilic Magnetospirillum strains in an oxygen-controlled fermentor. Appl Microbiol Biotechnol 61(5–6):536–544

    Article  CAS  Google Scholar 

  • Li X, Jiang W, Sun JB, Wang GL, Guan F, Li Y (2007a) Purified and sterilized magnetosomes from Magnetospirillum gryphiswaldense MSR-1 were not toxic to mouse fibroblasts in vitro. Lett Appl Microbiol 45(1):75–81

    Article  Google Scholar 

  • Li X, Wang B, Jin HL, Jiang W, Tian JS, Guan F, Li Y (2007b) Bacterial magnetic particles (BMPs)-PEI as a novel and efficient non-viral gene delivery system. J Gene Med 9(8):679–690

    Article  CAS  Google Scholar 

  • Liu Y, Li G, Guo F, Jiang W, Li Y, Li L (2010) Large-scale production of magnetosomes by chemostat culture of Magnetospirillum gryphiswaldense at high cell density. Microb Cell Fact 9(1):99

    Article  Google Scholar 

  • Matsunaga T, Tadokoro F, Nakamura N (1990) Mass culture of magnetic bacteria and their application to flow type immunoassays. IEEE T Magn 26(5):1557–1559

    Article  CAS  Google Scholar 

  • Matsunaga T, Kawasaki M, Yu X, Tsujimura N, Nakamura N (1996a) Chemiluminescence enzymes immunoassay using bacterial magnetic particles. Anal Chem 68(20):3551–3554

    Article  CAS  Google Scholar 

  • Matsunaga T, Tsujimura N, Kamiya S (1996b) Enhancement of magnetic particle production by nitrate and succinate fed-batch culture of Magnetospirillum sp. AMB-1. Biotechnol Tech 10(7):495–500

    Article  CAS  Google Scholar 

  • Matsunaga T, Tsujimura N, Okamura Y, Takeyama H (2000) Cloning and characterization of a gene, mpsA, encoding a protein associated with intracellular magnetic particles from Magnetospirillum sp. strain AMB-1. Biochem Biophys Res Commun 268(3):932–937

    Article  CAS  Google Scholar 

  • Matsunaga T, Arakaki A, Takahoko M (2002) Preparation of luciferase-bacterial magnetic particle complex by artificial integration of MagA-luciferase fusion protein into bacterial magnetic particle membrane. Biotechnol Bioeng 77:614–618

    Article  CAS  Google Scholar 

  • Matsunaga T, Takahashi M, Yoshino T, Kuhara M, Takeyama H (2006) Magnetic separation of CD14+ cells using antibody binding with protein A expressed on bacterial magnetic particles for generating dendritic cells. Biochem Biophys Res Commun 350(4):1019–1025

    Article  CAS  Google Scholar 

  • Nakamura N, Matsunaga T (1993) Highly sensitive detection of allergen using bacterial magnetic particles. Anal Chim Acta 281(3):585–589

    Article  CAS  Google Scholar 

  • Nakamura N, Hashimoto K, Matsunaga T (1991) Immunoassay method for the determination of immunoglobulin G using bacterial magnetic particles. Anal Chem 63(3):268–272

    Article  CAS  Google Scholar 

  • Okamura Y, Takeyama H, Matsunaga T (2000) Two-dimensional analysis of proteins specific to the bacterial magnetic particle membrane from Magnetospirillum sp. AMB-1. Appl Biochem Biotechnol 84–86:441–446

    Article  Google Scholar 

  • Okamura Y, Takeyama H, Matsunaga T (2001) A magnetosome-specific GTPase from the magnetic bacterium Magnetospirillum magneticum AMB-1. J Biol Chem 276(51):48183–48188

    Article  CAS  Google Scholar 

  • Okuda Y, Denda K, Fukumori Y (1996) Cloning and sequencing of a gene encoding a new member of the tetratricopeptide protein family from magnetosomes of Magnetospirillum magnetotacticum. Gene 171(1):99–102

    Article  CAS  Google Scholar 

  • Schellekens H, El-Gewely MR (2008) How to predict and prevent the immunogenicity of therapeutic proteins. Biotechnol Annu Rev 14:191–202

    Article  CAS  Google Scholar 

  • Schüler D, Baeuerlein E (1996) Iron-limited growth and kinetics of iron uptake in Magnetospirllum gryphiswaldense. Arch Microbiol 166(5):301–307

    Article  Google Scholar 

  • Schüler D, Baeuerlein E (1998) Dynamics of iron uptake and Fe3O4 mineralization during aerobic and microaerobic growth of Magnetospirillum gryphiswaldense. J Bacteriol 180(1):159–162

    Article  Google Scholar 

  • Sun JB, Duan JH, Dai SL, Ren J, Zhang YD, Tian JS, Li Y (2007) In vitro and in vivo antitumor effects of doxorubicin loaded with bacterial magnetosomes (DBMs) on H22 cells: the magnetic bio-nanoparticles as drug carriers. Cancer Lett 258:109–117

    Article  CAS  Google Scholar 

  • Sun JB, Duan JH, Dai SL, Ren J, Guo L, Jiang W, Li Y (2008a) Preparation and anti-tumor efficiency evaluation of doxorubicin-loaded bacterial magnetosomes: magnetic nanoparticles as drug carriers isolated from Magnetospirillum gryphiswaldense. Biotechnol Bioeng 101(6):1313–1320

    Article  CAS  Google Scholar 

  • Sun JB, Zhao F, Tang T, Jiang W, Tian JS, Li Y, Li JL (2008b) High-yield growth and magnetosome formation by Magnetospirillum gryphiswaldense MSR-1 in an oxygen-controlled fermentor supplied solely with air. Appl Microbiol Biotechnol 79(3):389–397

    Article  CAS  Google Scholar 

  • Switzer RC, Merrill CR, Shifrin S (1979) A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal Biochem 98:231–237

    Article  CAS  Google Scholar 

  • Tanaka M, Okamura Y, Arakaki A, Tanaka T, Takeyama H, Matsunaga T (2006) Origin of magnetosome membrane: proteomic analysis of magnetosome membrane and comparison with cytoplasmic membrane. Proteomics 6:5234–5247

    Article  CAS  Google Scholar 

  • Wacker R, Ceyhan B, Alhorn P, Schüler D, Lang C, Niemeyer CM (2007) Magneto Immuno-PCR: a novel immunoassay based on biogenic magnetosome nanoparticles. Biochem Biophys Res Commun 357:391–396

    Article  CAS  Google Scholar 

  • Wolff JA, Malone RW, Williams P, Chong W, Acsadi G, Jani A, Felgner PL (1990) Direct gene transfer into mouse muscle in vivo. Science 247(4949):1465–1468

    Article  CAS  Google Scholar 

  • Yang CD, Takeyama H, Matsunga T (2001a) Iron feeding optimization and plasmid stability in production of recombinant bacterial magnetic particles by Magnetospirillum magneticum AMB-1 in fed-batch culture. J Biosci Bioeng 91:213–216

    Article  CAS  Google Scholar 

  • Yang CD, Takeyama H, Tanaka T, Matsunaga T (2001b) Effects of growth medium composition, iron sources and atmospheric oxygen concentrations on production of luciferase-bacterial magnetic particle complex by a recombinant Magnetospirillum magneticum AMB-1. Enzyme Microb Technol 29:13–19

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by the High Technology Research and Development Program of China (Grant No. 2006AA02Z233 and 2007AA021805) and the National Natural Science Foundation of China (Grants No. 30870043). We would also like to thank Professor Song Tao for providing the magnet.

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Authors and Affiliations

  1. State Key Laboratories for Agro-biotechnology and College of Biological Sciences, China Agricultural University, Beijing, 100193, People’s Republic of China

    Fangfang Guo, Yang Liu, Yanping Chen, Tao Tang, Wei Jiang, Ying Li & Jilun Li

  2. France-China Bio-Mineralization and Nano-Structures Laboratory, Beijing, 100193, China

    Fangfang Guo, Yang Liu, Yanping Chen, Tao Tang, Wei Jiang, Ying Li & Jilun Li

Authors
  1. Fangfang Guo
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  2. Yang Liu
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  3. Yanping Chen
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  4. Tao Tang
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  5. Wei Jiang
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  6. Ying Li
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  7. Jilun Li
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Correspondence to Wei Jiang.

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Guo, F., Liu, Y., Chen, Y. et al. A novel rapid and continuous procedure for large-scale purification of magnetosomes from Magnetospirillum gryphiswaldense . Appl Microbiol Biotechnol 90, 1277–1283 (2011). https://doi.org/10.1007/s00253-011-3189-3

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  • DOI: https://doi.org/10.1007/s00253-011-3189-3

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