Impact of Glycerol as Carbon Source onto Specific Sugar and Inducer Uptake Rates and Inclusion Body Productivity in E. coli BL21(DE3)
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bioreactor Cultivations
2.2. Cultivation Scheme and qs Screening Procedure
2.3. Process Analytics
3. Results and Discussion
3.1. Mechanistic Correlations of Glycerol onto Specific Lactose Uptake Rate
- (i)
- Adaption phase: lactose gets transferred to alloactose and loads the induction (0–2 h in induction phase).
- (ii)
- Linear decrease of lactose as the system needs inducer for recombinant protein expression (2–5 h).
- (iii)
- Limitation of lactose in P strain: not sufficient inducer present, need for mixed feed system (5–7 h), no inducer limitation seen in NP strain, further decrease of inducer analogue to phase 2.
3.2. Productivity and Physiology Using Glycerol as Primary Carbon Source
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Amount of Glucose | Amount of Glycerol | |
---|---|---|
Preculture | 8.8 g/L | 8.9 g/L |
Batch-Media | 22 g/L | 23 g/L |
Feed | either 250 g/L or 300 g/L |
Cultivation System | qs,lac,max | KA | qs,C,crit | n | qs,lac,noglu | NRMSE |
---|---|---|---|---|---|---|
[g/g/h] | [g/g/h] | [g/g/h] | [-] | [g/g/h] | [%] | |
Glucose | 0.23 | 0.032 | 0.94 | 1.14 | 0.039 | 6.5 |
Glycerol | 0.23 | 0.053 | 0.74 | 0.74 | 0.051 | 2.6 |
Cultivation System | qs,lac,max [g/g/h] | KA [g/g/h] | qs,glu,crit [g/g/h] | n [-] | qs,lac,noglu [g/g/h] | NRMSE [%] |
---|---|---|---|---|---|---|
Glucose [NP] | 0.14 | 0.016 | 0.96 | 2.92 | 0.032 | 12.7 |
Glycerol [NP] | 0.10 | 0.13 | 0.78 | 0.90 | 0.029 | 9.7 |
Glucose | Glucose NP | Glycerol | Glycerol NP | ||||
---|---|---|---|---|---|---|---|
qs,C [g/g/h] | qCO2 [g/g/h] | qs,C [g/g/h] | qCO2 [g/g/h] | qs,C [g/g/h] | qCO2 [g/g/h] | qs,C [g/g/h] | qCO2 [g/g/h] |
0.036 | 2.15 ± 0.33 | 0.066 | 1.69 ± 0.25 | 0.022 | 2.91 ± 0.46 | 0.064 | 0.82 ± 0.09 |
0.116 | 3.12 ± 0.46 | 0.196 | 3.75 ± 0.44 | 0.054 | 4.41 ± 0.78 | 0.136 | 1.85 ± 0.21 |
0.197 | 3.98 ± 0.55 | 0.224 | 3.35 ± 0.42 | 0.093 | 3.88 ± 0.64 | 0.225 | 2.86 ± 0.31 |
0.286 | 5.72 ± 0.41 | 0.36 | 5.96 ± 0.26 | 0.159 | 3.12 ± 0.43 | 0.331 | 3.31 ± 0.22 |
0.403 | 6.42 ± 1.48 | 0.448 | 5.64 ± 0.47 | 0.199 | 4.14 ± 0.64 | 0.428 | 4.07 ± 0.51 |
0.544 | 7.30 ± 1.64 | 0.323 | 5.13 ± 0.48 | 0.603 | 1.75 ± 1.58 | ||
0.559 | 7.18 ± 2.10 |
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Kopp, J.; Slouka, C.; Ulonska, S.; Kager, J.; Fricke, J.; Spadiut, O.; Herwig, C. Impact of Glycerol as Carbon Source onto Specific Sugar and Inducer Uptake Rates and Inclusion Body Productivity in E. coli BL21(DE3). Bioengineering 2018, 5, 1. https://doi.org/10.3390/bioengineering5010001
Kopp J, Slouka C, Ulonska S, Kager J, Fricke J, Spadiut O, Herwig C. Impact of Glycerol as Carbon Source onto Specific Sugar and Inducer Uptake Rates and Inclusion Body Productivity in E. coli BL21(DE3). Bioengineering. 2018; 5(1):1. https://doi.org/10.3390/bioengineering5010001
Chicago/Turabian StyleKopp, Julian, Christoph Slouka, Sophia Ulonska, Julian Kager, Jens Fricke, Oliver Spadiut, and Christoph Herwig. 2018. "Impact of Glycerol as Carbon Source onto Specific Sugar and Inducer Uptake Rates and Inclusion Body Productivity in E. coli BL21(DE3)" Bioengineering 5, no. 1: 1. https://doi.org/10.3390/bioengineering5010001