Production of the soluble human Fas ligand by Dictyostelium discoideum cultivated on a synthetic medium.
Human Fas ligand (hFasL) is of considerable interest since it is a type II transmembrane glycoprotein that induces programmed cell death, or apoptosis. In this study Dictyostelium discoideum was used to produce a soluble form of the human Fas ligand. The recombinant cells were adapted to a modified synthetic FM medium, called SIH medium. Cells adapted to the SIH medium reached about 2 times higher cell densities and hFasL concentrations on this medium compared with cells growing on the standard complex medium HL-5C. Even higher values were achieved by a dissolved oxygen-controlled fed-batch cultivation in a conventional stirred bioreactor on SIH medium. Cell densities of up to 5.5 x 10(7) ml(-1) and a maximum hFasL concentration of 148 microgl(-1) were obtained. These results were further improved by means of continuous cultivation of D. discoideum in a bioreactor equipped with cell retention by microfiltration. At low space velocity very high cell densities of up to 2.4 x 10(8) ml(-1) and hFasL concentrations of up to 205 microgl(-1) were achieved.
Lu
Yinghua
Y
Department of Biochemistry, University of Groningen, Nijenborgh 4, NL-9747 AG Groningen, The Netherlands.
Knol
Jaco C
JC
Linskens
Maarten H K
MH
Friehs
Karl
K
Van Haastert
Peter J M
PJ
Flaschel
Erwin
E
D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism; D004023:Dictyostelium / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D053222:Fas Ligand Protein; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012995:Solubility
D003470:Culture Media; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins