Secretory Production of Recombinant Protein by a High Cell Density Culture of a Protease Negative Mutant Escherichia coli Strain

1999 ◽  
Vol 15 (2) ◽  
pp. 164-167 ◽  
Author(s):  
S.J. Park ◽  
G. Georgiou ◽  
S.Y. Lee
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Cell Density ◽  
High Cell Density ◽  
Coli Strain ◽  
High Cell Density Culture ◽  
High Cell ◽  
Secretory Production ◽  
Negative Mutant

scholarly journals Proteomic Profiling of Recombinant Escherichia coli in High-Cell-Density Fermentations for Improved Production of an Antibody Fragment Biopharmaceutical

2005 ◽  
Vol 71 (4) ◽  
pp. 1717-1728 ◽  
Author(s):  
Ilana S. Aldor ◽  
Denise C. Krawitz ◽  
William Forrest ◽  
Christina Chen ◽  
Julie C. Nishihara ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Cell Density ◽  
High Cell Density ◽  
Antibody Fragment ◽  
Culture Conditions ◽  
Test Method ◽  
Differentially Expressed ◽  
Soluble Form ◽  
High Cell

ABSTRACT By using two-dimensional polyacrylamide gel electrophoresis, a proteomic analysis over time was conducted with high-cell-density, industrial, phosphate-limited Escherichia coli fermentations at the 10-liter scale. During production, a recombinant, humanized antibody fragment was secreted and assembled in a soluble form in the periplasm. E. coli protein changes associated with culture conditions were distinguished from protein changes associated with heterologous protein expression. Protein spots were monitored quantitatively and qualitatively. Differentially expressed proteins were quantitatively assessed by using a t-test method with a 1% false discovery rate as a significance criterion. As determined by this criterion, 81 protein spots changed significantly between 14 and 72 h (final time) of the control fermentations (vector only). Qualitative (on-off) comparisons indicated that 20 more protein spots were present only at 14 or 72 h in the control fermentations. These changes reflected physiological responses to the culture conditions. In control and production fermentations at 72 h, 25 protein spots were significantly differentially expressed. In addition, 19 protein spots were present only in control or production fermentations at this time. The quantitative and qualitative changes were attributable to overexpression of recombinant protein. The physiological changes observed during the fermentations included the up-regulation of phosphate starvation proteins and the down-regulation of ribosomal proteins and nucleotide biosynthesis proteins. Synthesis of the stress protein phage shock protein A (PspA) was strongly correlated with synthesis of a recombinant product. This suggested that manipulation of PspA levels might improve the soluble recombinant protein yield in the periplasm for this bioprocess. Indeed, controlled coexpression of PspA during production led to a moderate, but statistically significant, improvement in the yield.

Expression, High Cell Density Culture and Purification of Recombinant EC-SOD in Escherichia coli

2010 ◽  
Vol 162 (6) ◽  
pp. 1585-1598 ◽  
Author(s):  
Young-Jin Son ◽  
Ji-Young Bae ◽  
Seon-Ha Chong ◽  
Hui Sun Lee ◽  
Sang Hyun Mo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
High Cell Density ◽  
High Cell Density Culture ◽  
High Cell
Sign in / Sign up

Export Citation Format

Share Document