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Semicontinuous system for the production of recombinant mCherry protein in Chlamydomonas reinhardtii.

  • Arias, Cesar Andres Diaz1
  • Matsudo, Marcelo Chuei2
  • Ferreira-Camargo, Livia Seno3
  • Molino, João Vitor Dutra1
  • Mayfield, Stephen Patrick4
  • de Carvalho, João Carlos Monteiro1
  • 1 Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil. , (Brazil)
  • 2 Institute of Natural Resources, Federal University of Itajubá, Itajubá, MG, Brazil. , (Brazil)
  • 3 Center for Natural and Human Sciences, Federal University of ABC, Santo André, SP, Brazil. , (Brazil)
  • 4 Department of Molecular Biology, and The California Center for Algae Biotechnology, University of California, San Diego, California, USA.
Published Article
Biotechnology Progress
Wiley (John Wiley & Sons)
Publication Date
Nov 09, 2020
DOI: 10.1002/btpr.3101
PMID: 33169497


Biotechnology advances have allowed bacteria, yeasts, plants, mammalian and insect cells to function as heterologous protein expression systems. Recently, microalgae have gained attention as an innovative platform for recombinant protein production, due to low culture media cost, compared to traditional systems, as well as the fact that microalgae such as Chlamydomonas reinhardtii are considered safe (GRAS) by the Food and Drug Administration (FDA). Previous studies showed that recombinant protein production in traditional platforms by semicontinuous process increased biomass and bio product productivity, when compared to batch process. As there is a lack of studies on semicontinuous process for recombinant protein production in microalgae, the production of recombinant mCherry fluorescent protein was evaluated by semicontinuous cultivation of Chlamydomonas reinhardtii in bubble column photobioreactor. This semicontinuous cultivation process was evaluated in the following conditions: 20%, 40%, and 60% culture portion withdrawal. The highest culture withdrawal percentage (60%) provided the best results, as an up to 161% increase in mCherry productivity (454.5 RFU h-1 - Relative Fluorescence Unit h-1 ), in comparison to batch cultivation (174.0 RFU h-1 ) of the same strain. All cultivations were carried out for 13 days, at pH 7, temperature 25°C and, by semicontinuous process, two culture withdrawals were taken during the cultivations. Throughout the production cycles, it was possible to obtain biomass concentration up to 1.36 g L-1 . © 2020 American Institute of Chemical Engineers.

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