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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. 1 Introduction Recombinant DNA technology is one of the pillars that have supported the progress of Biotechnology during the past 50 years.
Recombinant structural proteins and enzymes have direct applications as biopharmaceuticals (hormones, cytokines, growth factors, antibodies, etc.) and in technological processes that are central to industrial fields Spain phone number list such as food and feed, detergents, textile, paper, waste management, organic synthesis and biofuels (Kirk et al., 2002; Overton, 2014; Puetz and Wurm, 2019). The production of recombinant proteins is an expanding industry with increasing economic significance. Reports have estimated that the value for the global enzymes market in 2021 exceeded $6 billion, and a steady growth projection is forecast for the following years.
At a compound annual growth rate of 6.3%–6.8% between 2019 and 2027 (Sutay Kocabas and Grumet, 2019; BlueWeave Consulting and Research Pvt Ltd, 2021; de la Fuente et al., 2021; Espina et al., 2021; Mesbah, 2022). Concerning biopharmaceuticals, as of 2018, more than 300 protein-based drugs , and almost a third of all pharmaceuticals in development were protein-based (Walsh, 2018). The market of protein-based biopharmaceuticals has only continued to grow, with numerous new drugs being approved each year (Martins et al., 2022), and global sales reaching $90.53 billion in.
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