Substrate oscillations boost recombinant protein release from Escherichia coli

Jazini, M ; Sharif University of Technology

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  1. Type of Document: Article
  2. DOI: 10.1007/s00449-013-1059-3
  3. Abstract:
  4. Intracellular production of recombinant proteins in prokaryotes necessitates subsequent disruption of cells for protein recovery. Since the cell disruption and subsequent purification steps largely contribute to the total production cost, scalable tools for protein release into the extracellular space is of utmost importance. Although there are several ways for enhancing protein release, changing culture conditions is rather a simple and scalable approach compared to, for example, molecular cell design. This contribution aimed at quantitatively studying process technological means to boost protein release of a periplasmatic recombinant protein (alkaline phosphatase) from E. coli. Quantitative analysis of protein in independent bioreactor runs could demonstrate that a defined oscillatory feeding profile was found to improve protein release, about 60%, compared to the conventional constant feeding rate. The process technology included an oscillatory postinduction feed profile with the frequency of 4 min. The feed rate was oscillated triangularly between a maximum (1.3-fold of the maximum feed rate achieved at the end of the fed-batch phase) and a minimum (45% of the maximum). The significant improvement indicates the potential to maximize the production rate, while this oscillatory feed profile can be easily scaled to industrial processes. Moreover, quantitative analysis of the primary metabolism revealed that the carbon dioxide yield can be used to identify the preferred feeding profile. This approach is therefore in line with the initiative of process analytical technology for science-based process understanding in process development and process control strategies
  5. Keywords:
  6. Bioprocess technology ; Oscillatory feeding ; Protein release ; Substrate-related stress ; Carbon dioxide ; Drug products plants ; Escherichia coli ; Feeding ; Phosphatases ; Recombinant proteins ; ALkaline phosphatase ; Bioprocesses ; Intracellular production ; Process analytical technology ; Process control strategies ; Process understanding ; Total production cost ; Molecular biology ; Dissolved oxygen ; Recombinant protein ; Biological model ; Bioreactor ; Carbon evolution rate ; Cell density ; Flow rate ; Limit of detection ; Mathematical parameters ; Nonhuman ; Oscillation ; Oxygen uptake rate ; Polyacrylamide gel electrophoresis ; Priority journal ; Process technology ; Protein analysis ; Protein secretion ; Quantitative analysis ; Respiratory quotient ; Biological rhythm ; Genetics ; Growth, development and aging ; Secretion (process)
  7. Source: Bioprocess and Biosystems Engineering ; Volume 37, Issue 5 , May , 2014 , Pages 881-890 ; ISSN: 16157591
  8. URL: http://link.springer.com/article/10.1007%2Fs00449-013-1059-3