Development of redox potential-driven fermentation process for recombinant protein expression
- PMID: 33064228
- DOI: 10.1007/s10529-020-03030-9
Development of redox potential-driven fermentation process for recombinant protein expression
Abstract
Objectives: A redox potential-driven fermentation, maintaining dissolved oxygen at a prescribed level while simultaneously monitoring the changes of fermentation redox potential, was developed to guide the cultivation progress of recombinant protein expression.
Results: A recombinant E. coli harboring prolinase-expressing plasmid (pKK-PepR2) was cultivated using the developed process. Two distinct ORP valleys were noticeable based on recorded profile. The first ORP valley is equivalent to the timing for the addition of inducing agent, and the second ORP valley serves to guide the timing for cell harvesting. The final prolinase activity is 0.172 μmol/mg/min as compared to that of 0.154 μmol/mg/min where the optical density was employed to guide the timing of inducer addition and an empirically determined length of the cultivation.
Conclusion: The developed process can be further modified to become an automatic operation.
Keywords: Bioprocess development; Fermentation; Fermentation redox potential; Recombinant protein.
References
-
- Cirkovas A, Sereikaite J (2010) Increase in the solubility of recombinant mink growth hormone at low cultivation temperature of E. coli. Biotechnol Biotechnol Equip 24:2169–2171 - DOI
-
- Dahod SK (1982) Redox potential as a better substitute for dissolved oxygen in fermentation process control. Biotechnol Bioeng 24:2123–2125 - DOI
-
- Harms J, Wang X, Kim T, Yang X, Tathore A (2008) Defining process design space for biotech products: case study of Pichia pastoris fermentation. Biotechnol Prog 24:655–662 - DOI
-
- Huang Y, Tanaka T (2015) Characterization of two putative prolinases (PepR1 and PepR2) from Lactobacillus plantarum WCFS1: occurrence of two isozymes with structural similarity and different catalytic properties. Biochim Biophys Acta Proteins Proteom 1854:91–100 - DOI
-
- Lin YH, Chien WS, Duan KJ (2010) Correlations between reduction–oxidation potential profiles and growth patterns of Saccharomyces cerevisiae during very-high-gravity fermentation. Process Biochem 45:765–770 - DOI
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