Recovery of nuclease produced by Lactococcus lactis using expanded bed ion exchange chromatography.
Expanded bed-ionic exchange chromatography (EB-IEC) was used for the recovery and purification of recombinant staphylococcal nuclease secreted by Lactococcus lactis. At the end of the fermentation process, the nuclease activity reached 39 U ml(-1). The EB-IEC performances were firstly evaluated with clarified culture broth. The isocratic elution with 0.5 M NaCl led to approximately 80% of nuclease recovery. Proceeding with 3-fold bed expansion resulted in a reduction of the resin capacity by a factor of 32% compared to the process in a packed bed configuration. Simplification of the early purification steps was reached by loading immediately the unclarified culture broth previously diluted to reduce conductivity. Presence of Cells did not affect the chromatography performances resulting in 55-fold purification with the same yield.
Bodo
E
E
Meurice R&D asbl, Av. E. Gryzon 1, 1070, Brussels, Belgium.
Durieux
A
A
Saint-Hubert
C
C
Lavallée
R
R
Boufflette
J-M
JM
Simon
J-P
JP
D019149:Bioreactors / Q000382:microbiology*; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002852:Chromatography, Ion Exchange / Q000379:methods*; D013294:Lactococcus lactis / Q000502:physiology*; D008836:Micrococcal Nuclease / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; D008836:Micrococcal Nuclease