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A new generation of microbial expression hosts and tools for the production of biotherapeutics and high-value enzymes

Deliverables

Optimised redox-engineered strains

Optimised redoxengineered strains

IP protected by patents, publications including review written by ESRs

IP protected by patents publications including review written by ESRs

Supervisory Board of the network
Public engagement activities completed
List of common and critical parameters for specific productivity

List of common and critical parameters for specific productivity

Knowledge-based models of redox processes
Molecular models of redox processes
Local training completed
Overall recruitment

Overall recruitment; ESRs recruited & full report on recruitment process, including gender equality.

ESRs awarded PhDs

ESRs awarded PhDs date depends on local regulations

2nd generation strains with enhanced disulfide bond fidelity
Report on gender equality monitoring throughout project
Best strains for oxidative folding identified
prototype strains for enhanced disulphide bond formation
Training by research completed
Training of trainers completed
Project website live
Prototype strains with engineered stress responses
Strains with tested engineered capabilities for oxidative folding
Rotations and secondments completed
Report on outcome quality assurance throughout project
Minimised product heterogeneity
Recruitment symposium organized
Strains with tested robustness
Constructs for scFv, Fab and industrial enzymes
Optimised strains with minimal stress
ESR positions advertised
Live cell redox measurements
Network-wide training completed

Networkwide training completed

2nd generation strains with engineered stress responses
CyDisCo stress responses defined
Plan for future dual or joint doctoral degrees

Publications

Production of neutralizing antibody fragment variants in the cytoplasm of E. coli for rapid screening: SARS-CoV-2 a case study

Author(s): Tungekar Aatir A, Rosario Recacha & Lloyd W. Ruddock
Published in: Scientific Reports, 2023, ISSN 2045-2322
Publisher: Nature Publishing Group
DOI: 10.1038/s41598-023-31369-2

Yields and product comparison between Escherichia coli BL21 and W3110 in industrially relevant conditions: anti-c-Met scFv as a case study

Author(s): Klaudia Arauzo-Aguilera, Luisa Buscajoni, Karin Koch, Gary Thompson, Matthias Berkemeyer, Colin Robinson
Published in: Microbial Cell Factories, 2023, ISSN 1475-2859
Publisher: BioMed Central
DOI: 10.1186/s12934-023-02111-4

Microbial protein cell factories fight back?

Author(s): Lukas A. Rettenbacher; Klaudia Arauzo-Aguilera; Luisa Buscajoni; Angel Castillo-Corujo; Borja Ferrero-Bordera; Aliki Kostopoulou; Rafael Moran-Torres; David Núñez-Nepomuceno; Ayşegül Öktem; Arianna Palma; Beatrice Pisent; Martina Puricelli; Tobias Schilling; Aatir A. Tungekar; Jonathan Walgraeve; David Humphreys; Tobias von der Haar; Brigitte Gasser; Diethard Mattanovich; Lloyd Ruddock; Jan M
Published in: Trends in Biotechnology, Issue 40 (5), May 2022, 2022, Page(s) 576-590, ISSN 0167-7799
Publisher: Elsevier BV
DOI: 10.1016/j.tibtech.2021.10.003

Biochemical analysis of Komagataella phaffii oxidative folding proposes novel regulatory mechanisms of disulfide bond formation in yeast

Author(s): Palma A, Rettenbacher LA, Moilanen A, Saaranen M, Pacheco-Martinez C, Gasser B, Ruddock L
Published in: Sci Rep, 2023, ISSN 2045-2322
Publisher: Nature Publishing Group
DOI: 10.1038/s41598-023-41375-z

So you want to express your protein in Escherichia coli?

Author(s): Tungekar AA, Castillo-Corujo A, Ruddock LW
Published in: Essays Biochem, 2021, ISSN 0071-1365
Publisher: Portland Press, Ltd.
DOI: 10.1042/ebc20200170

Refolding in the modern biopharmaceutical industry

Author(s): Buscajoni L, Martinetz MC, Berkemeyer M, Brocard C
Published in: Biotechnol Adv, 2022, ISSN 0734-9750
Publisher: Elsevier BV
DOI: 10.1016/j.biotechadv.2022.108050

Enhancing bacterial fitness and recombinant enzyme yield by engineering the quality control protease HtrA of Bacillus subtilis

Author(s): Walgraeve J, Ferrero-Bordera B, Maaß S, Becher D, Schwerdtfeger R, van Dijl JM, Seefried M.
Published in: Microbiology Spectrum, 2023, ISSN 2165-0497
Publisher: American Society for Microbiology
DOI: 10.1128/spectrum.01608-23

Highly efficient export of a disulphide-bonded protein to the periplasm and medium by the Tat pathway using CyDisCo in Escherichia coli

Author(s): Klaudia Arauzo-Aguilera, Mirva J Saaranen, Colin Robinson and Lloyd W Ruddock
Published in: MicrobiogyOpen, 2022, ISSN 2045-8827
Publisher: John Wiley and Sons Inc.
DOI: 10.1002/mbo3.1350

A quantitative interpretation of oxidative protein folding activity in Escherichia coli

Author(s): Rettenbacher LA and von der Haar T
Published in: Microbial Cell Factories, 2022, ISSN 1475-2859
Publisher: BioMed Central
DOI: 10.1186/s12934-022-01982-3

Control of COVID-19 Outbreaks under Stochastic Community Dynamics, Bimodality, or Limited Vaccination

Author(s): Goldenbogen, B., Adler, S.O., Bodeit, O., Wodke, J.A.H., Escalera‐Fanjul, X., Korman, A., Krantz, M., Bonn, L., Morán‐Torres, R., Haffner, J.E.L.
Published in: Adv. Sci, 2022, ISSN 2198-3844
Publisher: Wiley-VCH
DOI: 10.1002/advs.202200088

Enhancing bacterial fitness and recombinant enzyme yield by engineering the quality control protease HtrA of Bacillus subtilis

Author(s): Öktem A, Núñez-Nepomuceno D, Ferrero-Bordera B, Walgraeve J, Seefried M, Gesell Salazar M, Steil L, Michalik S, Maaß S, Becher D, Mäder U, Völker U, van Dijl JM.
Published in: Microbiol Spectr., 2023, ISSN 2165-0497
Publisher: American Society for Microbiology
DOI: 10.1128/spectrum.01778-23

Efficient Production of Fc Fusion Proteins in the Cytoplasm of Escherichia coli: Dissecting and Mitigating Redox Heterogeneity

Author(s): Tungekar AA, Ruddock LW
Published in: Int J Mol Sci., 2022, ISSN 1422-0067
Publisher: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/ijms232314740

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