Growth Kinetics of Escherichia coli and Expression of a Recombinant Protein and Its Isoforms under Heat Shock Conditions


By jpeza - Posted on 09 Julio 2009

Fecha Publicación: 
1 Ene 1996
Nombre de Revista: 
Datos del paper
Autor Principal: 
Wen Ryan
*
Volumen: 
12
Issue: 
5
Página Inicial: 
596
Página Final: 
601
Abstract: 

Preinduction culture conditions were found to have significant impact on the expression

and post-translational modification of a recombinant human protein in Escherichia

coli under heat shock conditions (30 to 42 °C shift). Higher preinduction growth rates(

íg) favored better cell viability, greater cell mass yields, and increased cloned gene

expression during induction. Formation of recombinant protein isoforms (those

containing N-modified lysine residues) exhibited an increasing trend with increasing

íg. The different extents of post-translational modifications were suspected to be linked

to the different concentrations of certain heat shock protein chaperones resulting from

different íg. In view of the extensive involvement of E. coli heat shock proteins in

cellular activitiessincluding the synthesis, processing, modification, and degradationof proteins

sat elevated temperatures, it is believed that íg dictated the cellular

resources available for synthesizing the heat shock proteins required for cell survival,

which in turn determined the ability of the cells to respond to the heat shock. With

a higher íg, both the synthesis of host proteins (as indicated by cell growth and survival)

and the cloned gene expression were enhanced. The results demonstrate that there

exists an intermediate íg for optimum production of the unmodified foreign protein in

a heat shock environment. More importantly, they also illustrate the feasibility of

improving the recombinant protein homogeneity in fermentation, thereby facilitating

downstream processing.

Dirección del Autor: 

Amgen, Inc., Thousand Oaks, California 91320

Keywords: 
Escherichia coli ; fermentation processes
Coautores: 

Paul Collier, Lawrence Loredo, Joseph Pope, and Raj Sachdev

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