LABORATORY-SCALE PERMEABILIZATION OF ESCHERICHIA-COLI-CELLS FOR RECOVERY OF A SMALL RECOMBINANT PROTEIN - STAPHYLOKINASE

Citation
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
Citations number
13
Categorie Soggetti
Biothechnology & Applied Migrobiology
Journal title
ISSN journal
0178515X
Volume
17
Issue
1
Year of publication
1997
Pages
35 - 38
Database
ISI
SICI code
0178-515X(1997)17:1<35:LPOEFR>2.0.ZU;2-M
Abstract
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.