Effect of E. Coli endotoxin on mammalian cell growth and recombinant protein production

1990 ◽  
Vol 26 (12) ◽  
pp. 1121-1122 ◽  
Author(s):  
J. Epstein ◽  
C. E. Kelly ◽  
M. M. Lee ◽  
P. K. Donahoe
Keyword(s):  
Cell Growth ◽  
Recombinant Protein ◽  
Mammalian Cell ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
E Coli ◽  
Mammalian Cell Growth

scholarly journals Recombinant protein production provoked accumulation of ATP, fructose-1,6-bisphosphate and pyruvate in E. coli K12 strain TG1

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Jan Weber ◽  
Zhaopeng Li ◽  
Ursula Rinas
Keyword(s):  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System ◽  
Limiting Factors ◽  
Expression Vectors ◽  
Metabolic Enzyme ◽  
E Coli ◽  
Metabolic Burden ◽  
Fructose 1,6 Bisphosphate

Abstract Background Recently it was shown that production of recombinant proteins in E. coli BL21(DE3) using pET based expression vectors leads to metabolic stress comparable to a carbon overfeeding response. Opposite to original expectations generation of energy as well as catabolic provision of precursor metabolites were excluded as limiting factors for growth and protein production. On the contrary, accumulation of ATP and precursor metabolites revealed their ample formation but insufficient withdrawal as a result of protein production mediated constraints in anabolic pathways. Thus, not limitation but excess of energy and precursor metabolites were identified as being connected to the protein production associated metabolic burden. Results Here we show that the protein production associated accumulation of energy and catabolic precursor metabolites is not unique to E. coli BL21(DE3) but also occurs in E. coli K12. Most notably, it was demonstrated that the IPTG-induced production of hFGF-2 using a tac-promoter based expression vector in the E. coli K12 strain TG1 was leading to persistent accumulation of key regulatory molecules such as ATP, fructose-1,6-bisphosphate and pyruvate. Conclusions Excessive energy generation, respectively, accumulation of ATP during recombinant protein production is not unique to the BL21(DE3)/T7 promoter based expression system but also observed in the E. coli K12 strain TG1 using another promoter/vector combination. These findings confirm that energy is not a limiting factor for recombinant protein production. Moreover, the data also show that an accelerated glycolytic pathway flux aggravates the protein production associated “metabolic burden”. Under conditions of compromised anabolic capacities cells are not able to reorganize their metabolic enzyme repertoire as required for reduced carbon processing.

Reduction of Acetate and Lactate Contributed to Enhancement of a Recombinant Protein Production in E. coli BL21

2015 ◽  
Vol 25 (7) ◽  
pp. 1093-1100 ◽  
Author(s):  
Tae-Su Kim ◽  
Hyung-Moo Jung ◽  
Sang-Yong Kim ◽  
Liaoyuan Zhang ◽  
Jinglin Li ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
E Coli

scholarly journals Decoupling of recombinant protein production from Escherichia coli cell growth enhances functional expression of plant Leloir glycosyltransferases

2019 ◽  
Vol 116 (6) ◽  
pp. 1259-1268 ◽  
Author(s):  
Martin Lemmerer ◽  
Juergen Mairhofer ◽  
Alexander Lepak ◽  
Karin Longus ◽  
Rainer Hahn ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Growth ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Functional Expression ◽  
Coli Cell
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