title: Integrated bioprocess for the production and purification of recombinant proteins by affinity chromatography in Escherichia coli.
abstract: In order to improve the effectiveness of the production of recombinant proteins in E. coli, integrated fermentation processes were developed. Therefore, expression vectors were constructed containing a strongly expressed gene for a beta-glucanase fused with a metal-chelating affinity tag and a leader peptide for directing the fusion protein into the periplasmic space. Its export into the medium was achieved by means of co-expression of a bacteriocin-release protein, the Kil protein from pColE1. Bioreactors were modified so that special devices containing metal chelate pentadentate chelator PDC resins were located within the bioreactor. Using the bioreactor with an internal device the Zn2+-PDC had a 4.3-fold higher binding capacity than metal-free PDC (12.3 and 2.6 kU ml(-1) PDC, respectively. Using the bioreactor with charged PDC in an external circuit revealed even higher beta-glucanase concentration (65.6 kU ml(-1)), i.e. 1.5-fold compared to the internal adsorbent system.
authors.affiliation: Bioprocess Development Department, Mubarak City for Scientific Research and Technology Applications, Genetic Engineering and Biotechnology Research Institute (GEBRI), New Borg El-Arab, Alexandria 21934, Egypt. ube@fermtech.techfak.uni-bielefeld.de
authors.forename: Usama
authors.initials: U
authors.lastname: Beshay
authors.forename: Gerhard
authors.initials: G
authors.lastname: Miksch
authors.forename: Karl
authors.initials: K
authors.lastname: Friehs
authors.forename: Erwin
authors.initials: E
authors.lastname: Flaschel
mesh_terms: D001483:Base Sequence; D018533:Biomass; D019149:Bioreactors; D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D004247:DNA