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Long-term production of soluble human Fas ligand through immobilization of Dictyostelium discoideum in a fibrous bed bioreactor

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Abstract

The production of recombinant glycoproteins in Dictyostelium discoideum by conventional cell culture methods was limited by low cell density as well as low growth rate. In this work, cotton towel with a good adsorption capability for D. discoideum cells was used as the immobilization matrix in an external fibrous bed bioreactor (FBB) system. With batch cultures in the FBB, the concentration of immobilized cells in the cotton fiber carrier increased to 1.37 × 108 cells per milliliter after 110-h cultivation, which was about tenfold higher than the maximal cell density in the conventional free-cell culture. Correspondingly, a high concentration of soluble human Fas ligand (hFasL; 173.7 μg l−1) was achieved with a high productivity (23 μg l−1 h−1). The FBB system also maintained a high density of viable cells for hFasL production during repeated-batch cultures, achieving a productivity of 9∼10 μg l−1 h−1 in all three batches studied during 15 days. The repeated-batch culture using immobilized cells of D. discoideum in the FBB system thus provides a good method for long-term and high-level production of hFasL.

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Reference

  1. Beshay U, Friehs K, Azzam AEM, Flaschel E (2003a) Analysis of the behavior of Dictyostelium discoideum in immobilized state by means of continuous cultivation. Bioproc Biosyst Eng 26:117–122

  2. Beshay U, Friehs K, Azzam AEM, Flaschel E (2003b) Cultivation of Dictyostelium discoideum in immobilized form by colonization of porous supports. Process Biochem 38:1521–1529

  3. Chen J, Lu YH, Xu ZN, Cen PL, Fang XM (2007) Efficient expression and primary purification of 6-his tagged human Fas ligand in Dictyostelium discoideum. Biotechnol Lett 29:859–863

  4. Glenn D, Williams KL (1988) Dictyostelium discoideum: its future in biotechnology. Austr J Biotech 1:46–51

  5. Huang Y, Yang ST (1998) Acetate production from whey lactose using co-immobilized cells of homolactic and homoacetic bacteria in a fibrous-bed bioreactor. Biotechnol Bioeng 60:498–507

  6. Jung E, Williams KL (1997) The production of recombinant glycoproteins with special reference to simple eukaryotes including Dictyostelium discoideum. Biotechnol Appl Bioc 25:3–8

  7. Looby D, Griffiths B (1990) Immobilization of animal cells in porous carrier culture. Trends Biotechnol 8:204–209

  8. Lu YH, Knol JC, Linskens MHK, Friehs K, Van Haastert PJM, Flaschel E (2004a) Production of the soluble human Fas ligand by Dictyostelium discoideum cultivated on a synthetic medium. J Biotechnol 108:243–251

  9. Lu YH, Knol JC, Linskens MHK, Friehs K, Van Haastert PJM, Flaschel E (2004b) Cultivation of immobilized Dictyostelium discoideum for the production of soluble human Fas ligand. Appl Microbiol Biot 65:547–552

  10. Lu YH, Wang Y, Wu XX, Xu ZN, He N, Chen J (2008) Influence of medium components on growth kinetics of Dictyostelium discoideum. World J Microbiol Biotech 24:491–499

  11. Rathi A, Kayman SC, Clarke M (1991) Induction of gene expression in Dictyostelium by pre-starvation factor, a factor secreted by growing cells. Dev Genet 12:82–87

  12. Silva EM, Yang ST (1995) Kinetics and stability of a fibrous-bed bioreactor for continuous production of lactic acid from unsupplemented acid whey. J Biotechnol 41:59–70

  13. Slade MB, Emslie KR, Williams KL (1997) Expression of recombinant glycoproteins in the simple eukaryote Dictyostelium discoideum. Biotechnol Genet Eng Rev 14:1–35

  14. Sussman RR, Sussman M (1967) Cultivation of Dictyostelium discoideum in axenic medium. Biochem Biophy Res Co 29:53–55

  15. Tanaka M, Suda T, Takahashi T, Nagata S (1995) Expression of the functional soluble form of human Fas ligand in activated lymphocytes. Embo J 14:1129–1135

  16. Varki A, Cummings R, Esko J, Freeze H, Hart G, Marth J, Chrispeels M, Hindsgaul O, Paulson JC, Lowe J, Manzi A, Powell L, Van Halbeek H (1999) Essentials of glycobiology. Cold Spring Harbor, New York

  17. Watts DJ, Ashworth JM (1970) Growth of myxamoebae of the cellular slime mould Dictyostelium discoidium in axinic culture. Biochem J 119:171–174

  18. Xu ZN, Yang ST (2007) Production of mycophenolic acid by immobilized cells of Penicillium brevicompactum in a rotating fibrous bed bioreactor. Enz Microb Technol 40(4):623–628

  19. Yarger J, Stults K, Soll DR (1974) Observations on the growth of Dictyostelium discoideum in axenic medium: evidence for an extracellular growth inhibitor synthesized by stationary phase cells. J Cell Sci 14:681–690

  20. Yang ST, Zhu H, Li Y, Hong G (1994) Continuous propionate production from whey permeate using a novel fibrous bed bioreactor. Biotechnol Bioeng 43:1124–1130

  21. Zhu H, Yang ST (2004) Long-term continuous production of monoclonal antibody by hybridoma cells immobilized in a fibrous-bed bioreactor. Cytotechnology 44:1–14

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Acknowledgement

This work was financially supported by the National Natural Science Foundation of China (nos. 20736008, 30370039, and 20676115), the National Basic Research Program of China (2007CB707805), and the Ministry of Science and Technology, China.

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Correspondence to Zhinan Xu.

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Chen, J., Chen, H., Zhu, X. et al. Long-term production of soluble human Fas ligand through immobilization of Dictyostelium discoideum in a fibrous bed bioreactor. Appl Microbiol Biotechnol 82, 241–248 (2009). https://doi.org/10.1007/s00253-008-1769-7

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Keywords

  • Dictyostelium discoideum
  • Immobilization
  • Soluble human Fas ligand
  • Fibrous bed bioreactor