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.

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Fig. 1

Abbreviations

DO:

Dissolved oxygen

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

OD:

Optical density

ORP:

Fermentation redox potential

References

  1. 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

    CAS  Article  Google Scholar 

  2. Dahod SK (1982) Redox potential as a better substitute for dissolved oxygen in fermentation process control. Biotechnol Bioeng 24:2123–2125

    CAS  Article  Google Scholar 

  3. 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

    CAS  Article  Google Scholar 

  4. 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

    CAS  Article  Google Scholar 

  5. 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

    CAS  Article  Google Scholar 

  6. Liu CG, Qin JC, Lin YH (2017) Fermentation processes. In: Jozala A (ed) Fermentation and redox potential. IntechOpen, London, pp 23–42

    Google Scholar 

  7. Ou J, Wang L, Ding X, Du J, Zhang Y, Chen H, Xu A (2004) Stationary phase protein overproduction is a fundamental capability of Escherichia coli. Biochem Biophys Res Res 314:174–180

    CAS  Article  Google Scholar 

  8. San-Miguel T, Pérez-Bermúdez P, Gavidia I (2013) Production of soluble eukaryotic recombinant proteins in E. coli is favoured in early log-phase cultures induced at low temperature. Springerplus 2:1–4

    Article  Google Scholar 

  9. Stevenson K, McVey A, Clark I, Swain P, Pilizota T (2016) General calibration of microbial growth in microplate readers. Sci Rep 6:38828

    CAS  Article  Google Scholar 

Download references

Funding

The authors acknowledge the financial support from the Natural Sciences and Engineering Research Council of Canada.

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Correspondence to Yen-Han Lin.

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The authors declare that there is no conflict of interest.

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Guo, J., Wu, Y., Tanaka, T. et al. Development of redox potential-driven fermentation process for recombinant protein expression. Biotechnol Lett 43, 99–103 (2021). https://doi.org/10.1007/s10529-020-03030-9

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Keywords

  • Fermentation redox potential
  • Recombinant protein
  • Fermentation
  • Bioprocess development