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Masters Thesis | Biochemistry | India | Volume 11 Issue 7, July 2022 | Popularity: 4.8 / 10
Effect of Carbon Concentration in Batch Media on Up-Scaling Biomass Production
Nandini Banerjee
Abstract: Now a days recombinant bacteria (commonly used strain E. Coli BL21) is commonly used as a host and expression strain for protein expression and purification. When large quantities of fully deuterated proteins are required, E. coli is often grown in minimal media or customize chemical defined culture media with glucose or dextrose as the carbon source because these are less expensive. Despite the widespread use of BL21, we want to study the effect of different carbon sources concentration on BL21 growth. In this study, we assessed the growth behavior of E. coliBL21 in chemical defined media with different gluconeogenic carbon sources concentration. Though BL21 grew reasonably well on glycerol and pyruvate, it had a prolonged lag-phase on succinate (20 h), acetate (10 h), and fumarate (20 h), attributed to the physiological adaptation of E. coli cells grew well on all the substrates. We also examined the growth of E. coli BL21 in bioreactor to optimized bacterial batch growth using chemical defined culture media. The bioreactors is used to carefully control temperature, pH, and dissolved oxygen concentrations, in particular, makes them essential to efficient large-scale growth and expression of fermentation products. Scale-up from shake flasks to bioreactor has been hampered by the lack of knowledge, so before start commercial product, I have run bioreactor only for biomass production. Escherichia coli has a proven track record for successful production of anything from small molecules like organic acids to large therapeutic proteins, and has thus important applications in both R&D and commercial production. E. Coli is a very fast growing bacterial strain, for optimization study it is an ideal bacterial strain. We used chemical-defined media for the growth of E.coli rapidly in large quantities. In this experiment I used Carbon sources which is provide energy and major building blocks elements for the microorganism for growth and their choice is very critical for the production of high levels of antibiotics. For any lab-based experiment shake flask bacterial growth is good but when thinking about a large amount of production, means thinking about scale-up, then bioreactor is the only option. So that optimization study about up-scaling from shake flask to bench-top bioreactor is necessary.
Keywords: Escherichia coli, wet cell mass, dry cell mass, bioprocessing, up-scale bench bioreactor, shake flask bioreactor, bacterial growth curve
Edition: Volume 11 Issue 7, July 2022
Pages: 1043 - 1055
DOI: https://www.doi.org/10.21275/SR22616093808
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