High-level production of recombinant human IFN-α2a with co-expression of tRNAArg(AGG/AGA) in high-cell-density cultures ofEscherichia coli

2001 ◽  
Vol 6 (4) ◽  
pp. 301-305 ◽  
Author(s):  
Chul Soo Shin ◽  
Min Seon Hong ◽  
Hang-Cheol Shin ◽  
Jeewon Lee
Keyword(s):  
Cell Density ◽  
High Cell Density ◽  
Ofescherichia Coli ◽  
High Cell ◽  
High Cell Density Cultures ◽  
High Level ◽  
Level Production

scholarly journals The Presence of N-Terminal Secretion Signal Sequences Leads to Strong Stimulation of the Total Expression Levels of Three Tested Medically Important Proteins during High-Cell-Density Cultivations of Escherichia coli

2006 ◽  
Vol 73 (3) ◽  
pp. 906-912 ◽  
Author(s):  
H. Sletta ◽  
A. T�ndervik ◽  
S. Hakv�g ◽  
T. E. Vee Aune ◽  
A. Nedal ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
High Cell Density ◽  
Signal Sequence ◽  
High Cell ◽  
Signal Sequences ◽  
Expression Levels ◽  
Gm Csf ◽  
High Level ◽  
Level Production

ABSTRACT Genetic optimizations to achieve high-level production of three different proteins of medical importance for humans, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon alpha 2b (IFN-α2b), and single-chain antibody variable fragment (scFv-phOx), were investigated during high-cell-density cultivations of Escherichia coli. All three proteins were poorly expressed when put under control of the strong Pm/xylS promoter/regulator system, but high volumetric yields of GM-CSF and scFv-phOx (up to 1.7 and 2.3 g/liter, respectively) were achieved when the respective genes were fused to a translocation signal sequence. The choice of signal sequence, pelB, ompA, or synthetic signal sequence CSP, displayed a high and specific impact on the total expression levels for these two proteins. Data obtained by quantitative PCR confirmed relatively high in vivo transcript levels without using a fused signal sequence, suggesting that the signal sequences mainly stimulate translation. IFN-α2b expression remained poor even when fused to a signal sequence, and an alternative IFN-α2b coding sequence that was optimized for effective expression in Escherichia coli was therefore synthesized. The total expression level of this optimized gene remained low, while high-level production (0.6 g/liter) was achieved when the gene was fused to a signal sequence. Together, our results demonstrate a critical role of signal sequences for achieving industrial level expression of three human proteins in E. coli under the conditions tested, and this effect has to our knowledge not previously been systematically investigated.

Change of extracellular cAMP concentration is a sensitive reporter for bacterial fitness in high-cell-density cultures ofEscherichia coli

2004 ◽  
Vol 87 (5) ◽  
pp. 602-613 ◽  
Author(s):  
Hongying Lin ◽  
Frank Hoffmann ◽  
Aleksei Rozkov ◽  
Sven-Olof Enfors ◽  
Ursula Rinas ◽  
...  
Keyword(s):  
Cell Density ◽  
High Cell Density ◽  
Ofescherichia Coli ◽  
High Cell ◽  
Camp Concentration ◽  
Bacterial Fitness ◽  
High Cell Density Cultures ◽  
Extracellular Camp

Combined transcriptome and proteome analysis ofEscherichia coli during high cell density culture

2003 ◽  
Vol 81 (7) ◽  
pp. 753-767 ◽  
Author(s):  
Sung Ho Yoon ◽  
Mee-Jung Han ◽  
Sang Yup Lee ◽  
Ki Jun Jeong ◽  
Jong-Shin Yoo
Keyword(s):  
Cell Density ◽  
High Cell Density ◽  
Proteome Analysis ◽  
Ofescherichia Coli ◽  
High Cell Density Culture ◽  
High Cell
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