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University of Groningen

A Tale of Two Cell Factories Neef, Jolanda

DOI:

10.33612/diss.99279788

IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below.

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Publication date: 2019

Link to publication in University of Groningen/UMCG research database

Citation for published version (APA):

Neef, J. (2019). A Tale of Two Cell Factories: Heterologous protein secretion in Bacillus subtilis and Lactococcus lactis. University of Groningen. https://doi.org/10.33612/diss.99279788

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A Tale of Two Cell Factories 

 

Heterologous protein secretion in  

Bacillus subtilis and Lactococcus lactis 

 

 

 

 

 

 

 

 

Jolanda Neef 

 

 

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The  work  described  in  this  thesis  was  performed  in  the  Laboratory  of  Molecular  Bacteriology,  Department of Medical Microbiology of the University of Groningen and the University Medical Center  of Groningen (The Netherlands).  

The studies presented in this thesis were financially supported by the Top Institute Pharma (Project  T4‐213 and T4‐502) and Genencor‐DuPont (Palo Alto, CA, USA). 

Publication  of  this  thesis  was  partly  supported  by  the  Graduate  School  of  Medical  Sciences  of  the  University of Groningen.                                    

A  Tale  of  Two  Cell  Factories  –  Heterologous  protein  secretion  in  Bacillus  subtilis  and  Lactococcus 

lactis  Dissertation of the University of Groningen  ISBN: 978‐94‐034‐2101‐8 (printed book)   ISBN: 978‐94‐034‐2100‐1 (digital)  Cover: Abstract interpretation of the Two Cell Factories, Bacillus subtilis and Lactococcus lactis  Cover design: Amber Nieuwenweg and Jolanda Neef  Printed by: Ipskamp Printing, Enschede  Copyright © Jolanda Neef, 2019 

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A Tale of Two Cell Factories 

Heterologous protein secretion in Bacillus subtilis and Lactococcus lactis 

 

 

PhD thesis 

 

 

to obtain the degree of PhD at the   University of Groningen  on the authority of the  Rector Magnificus Prof. C. Wijmenga  and in accordance with   the decision of the College of Deans    This thesis will be defended in public on    Wednesday 6 November 2019 at 12:45 hours    by    Jolanda Neef    born on 14 August 1978  in Bedum

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Supervisor  Prof. J. M. van Dijl    Co‐supervisor  Dr. G. Buist    Assessment committee    Prof. J. Kok  Prof. D.J. Scheffers  Prof. D. van Sinderen 

 

 

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Paranymphs  Dr. Sjouke Piersma  Drs. Rocío Aguilar Suárez                      Voor Joep en Nienke 

 

 

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Table of content 

 

Chapter 1  General introduction and scope of this thesis        9      Manuscript to be submitted        Chapter 2  Efficient production of secreted staphylococcal antigens in a     43  non‐lysing and proteolytically reduced Lactococcus lactis strain  Published in Appl Microbiol Biotechnol. 2014, 98: 10131‐41    Chapter 3  Versatile vector suite for the extracytoplasmic production and    67      purification of heterologous His‐tagged proteins in Lactococcus      lactis      Published in Appl Microbiol Biotechnol. 2015, 99: 9037‐48    Chapter 4  A Lactococcus lactis expression vector set with multiple affinity     89  tags to facilitate isolation and direct labeling of heterologous secreted  proteins  Published in Appl Microbiol Biotechnol. 2017, 101: 8139–49    Chapter 5  Human antibody responses against non‐covalently cell wall‐bound   107  Staphylococcus aureus proteins    Published in Sci Rep. 2018, 8: 3234    Chapter 6  Intramembrane protease RasP boosts protein production in Bacillus  131    Published in Microb Cell Fact. 2017, 16: 57    Chapter 7  Relative contributions of individual Sec pathway components to high‐  149  level enzyme secretion by Bacillus subtilis  Manuscript to be submitted    Chapter 8  General summary and discussion      169    Chapter 9  Nederlandse samenvatting (voor de leek)      179    Chapter 10  Dankwoord – Acknowledgements      189      Curriculum Vitae (NL)       197      Curriculum Vitae (ENG)      198      List of publications      199   

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