I. Gehmlich et al., LABORATORY-SCALE PERMEABILIZATION OF ESCHERICHIA-COLI-CELLS FOR RECOVERY OF A SMALL RECOMBINANT PROTEIN - STAPHYLOKINASE, Bioprocess engineering, 17(1), 1997, pp. 35-38
The recovery of recombinant proteins includes a purification process t
hat has to be compressed to a minimum of steps in order to get high yi
elds with a low cost expenditure. A selective liberation of recombinan
t proteins by cell permeabilization leads to both a high product purit
y just in the beginning of the recovery process and to a simplificatio
n of the cell residue separation compared to the mechanical cell disru
ption. In case of the purification of the bacterial plasminogen activa
tor Staphylokinase from E. coli cells, yields of 82% with a purity of
46% were attained by utilization of permeabilization by biomass freezi
ng, resuspension in a Tris/EDTA-buffer and following micro-diafiltrati
on. A recovery process without interrupt ion (freezing) is possible du
e to the addition of guanidine-HCl and Triton X100 to the buffer. Thes
e methods were developed on a laboratory-scale.