Biomanufacturing Potential of Streamlined Cells

Biotechnology and Bioengineering, vol. 122, pp. 3309–3318

Abstract

A series of Escherichia coli streamlined strains was developed by removing the expression of genes encoding extracellular structures and unessential enzymes. The streamlined strains exhibited improved metabolic performance, including lower overflow metabolism and ATP maintenance coefficient, as well as a higher growth rate, compared to their parental strain. The intracellular levels of ATP were monitored using a genetic sensor, showing the improved resource stewardship of the streamlined cells. The streamlined strains were tested as cell factories to produce plasmid DNA (pDNA) in batch cultures, exhibiting a 23% increase in the specific pDNA production rate, compared to the parental strain. Recombinant protein expression was evaluated in microbioreactors in batch and fed-batch modes. In batch mode, recombinant protein yield from biomass was up to 82% higher in the streamlined strains than in the parental strain. Furthermore, in fed-batch mode, the recombinant protein yield was 79% greater in the streamlined cells compared to the parental strain. Our results show the benefits of reducing cellular complexity on the biomanufacturing of pDNA and recombinant proteins in culture schemes typical of industrial settings.

Authors 10

1. Alvaro R. Lara corresponding

   Aarhus University

   Affiliation as printed

   Department of Biological and Chemical Engineering Aarhus University Aarhus Denmark

   Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark

2. Marie B. Andersen

   Aarhus University

   Affiliation as printed

   Department of Biological and Chemical Engineering Aarhus University Aarhus Denmark

   Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark

3. Kathrine Gravlund Fønss

   Aarhus University

   Affiliation as printed

   Department of Biological and Chemical Engineering Aarhus University Aarhus Denmark

   Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark

4. Marta Irla

   Aarhus University

   Affiliation as printed

   Department of Biological and Chemical Engineering Aarhus University Aarhus Denmark

   Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark

5. Hilal Taymaz‐Nikerel

   Boğaziçi University

   Affiliation as printed

   Department of Chemical Engineering Boğaziçi University Istanbul Turkey

6. Luz María Martínez

   Universidad Nacional Autónoma de México

   Affiliation as printed

   Instituto de Biotecnología Universidad Nacional Autónoma de México Cuernavaca México

7. Max Daniel Dicke Aachen Chair of Biochemical Engineering (AVT.BioVT)

   RWTH Aachen University

   Affiliation as printed

   Chair of Biochemical Engineering (AVT.BioVT) RWTH Aachen University Aachen Germany

   Chair of Biochemical Engineering (AVT.BioVT), RWTH Aachen University, Aachen, Germany

8. Marcel Mann Aachen Chair of Biochemical Engineering (AVT.BioVT)

   RWTH Aachen University

   Affiliation as printed

   Chair of Biochemical Engineering (AVT.BioVT) RWTH Aachen University Aachen Germany

   Chair of Biochemical Engineering (AVT.BioVT), RWTH Aachen University, Aachen, Germany

9. Jørgen Barsett Magnus Aachen Chair of Biochemical Engineering (AVT.BioVT)

   RWTH Aachen University

   Affiliation as printed

   Chair of Biochemical Engineering (AVT.BioVT) RWTH Aachen University Aachen Germany

   Chair of Biochemical Engineering (AVT.BioVT), RWTH Aachen University, Aachen, Germany

10. Guillermo Gosset

    Universidad Nacional Autónoma de México

    Affiliation as printed

    Instituto de Biotecnología Universidad Nacional Autónoma de México Cuernavaca México