Central carbon metabolite profiling reveals vector-associated differences in the recombinant protein production host Escherichia coli BL21
Garcia Calvo, Laura; Rane, Divyata Vilas; Everson, Nikalet; Humlebrekk, Sigurd Tømmerberg; Mathiassen, Lise Femanger; Mæhlum, Astfrid Helene Morka; Malmo, Jostein; Bruheim, Per
Peer reviewed, Journal article
Published version
View/ Open
Date
2023Metadata
Show full item recordCollections
Original version
10.3389/fceng.2023.1142226Abstract
The Gram-negative bacterium Escherichia coli is the most widely used host for recombinant protein production, both as an industrial expression platform and as a model system at laboratory scale. The recombinant protein production industry generates proteins with direct applications as biopharmaceuticals and in technological processes central to a plethora of fields. Despite the increasing economic significance of recombinant protein production, and the importance of E. coli as an expression platform and model organism, only few studies have focused on the central carbon metabolic landscape of E. coli during high-level recombinant protein production. In the present work, we applied four targeted CapIC- and LC-MS/MS methods, covering over 60 metabolites, to perform an in-depth metabolite profiling of the effects of high-level recombinant protein production in strains derived from E. coli BL21, carrying XylS/Pm vectors with different characteristics. The mass-spectrometric central carbon metabolite profiling was complemented with the study of growth kinetics and protein production in batch bioreactors. Our work shows the robustness in E. coli central carbon metabolism when introducing increased plasmid copy number, as well as the greater importance of induction of recombinant protein production as a metabolic challenge, especially when strong promoters are used. Central carbon metabolite profiling reveals vector-associated differences in the recombinant protein production host Escherichia coli BL21