Protein secretion and disulfide bond handling in Bacillus subtilis Kouwen, Roelof Hendrik Matthijs

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1 Protein secretion and disulfide bond handling in Bacillus subtilis Kouwen, Roelof Hendrik Matthijs 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. Document Version Publisher's PDF, also known as Version of record Publication date: 2009 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Kouwen, R. H. M. (2009). Protein secretion and disulfide bond handling in Bacillus subtilis Groningen: s.n. Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date:

2 Protein secretion and disulfide bond handling in Bacillus subtilis

3 ISBN (printed version) (digital version) Printing Printed by PrintPartners Ipskamp, Enschede, the Netherlands Copyright All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means without the permission of the author and the publisher holding the copyright of the published articles. Cover The figure on the cover, showing two dimerized B. subtilis TrxA subunits, is a modified version of figure 3A in chapter 6 of this thesis. Original image made by Juni Andréll. Printed with permission of the author

4 RIJKSUNIVERSITEIT GRONINGEN Protein secretion and disulfide bond handling in Bacillus subtilis Proefschrift ter verkrijging van het doctoraat in de Medische Wetenschappen aan de Rijksuniversiteit Groningen op gezag van de Rector Magnificus, Dr. F. Zwarts, in het openbaar te verdedigen op woensdag 7 januari 2009 om uur door Roelof Hendrik Matthijs Kouwen geboren op 7 november 1979 te Groningen

5 Promotor: Copromotor: Prof. Dr. J.M. van Dijl Dr. J.-Y.F. Dubois Beoordelingscommissie: Prof. Dr. H.J. Busscher Prof. Dr. C.R. Harwood Prof. Dr. A.J.M. Driessen

6 Paranimfen: Mark J.J.B. Sibbald René van der Ploeg Our doubts are traitors, And make us lose the good we oft might win, By fearing to attempt. William Shakespeare, Measure for Measure

7 The studies described in this thesis were performed at the Faculty of Medical Sciences, Department of Medical Microbiology, section Molecular Bacteriology of the University Medical Center Groningen and University of Groningen within the Graduate School for Drug Exploration GUIDE. The research described in this thesis was supported through CEU project LSHG-CT (BACELL Health). Publication of this thesis was financially supported by Novozymes A/S, Genencor International B.V., DSM Nutritional Products and the Graduate School for Drug Exploration GUIDE. Their contributions are greatly appeciated.

8 Contents Chapter 1 Introduction and scope of this thesis 9 Chapter 2 Chapter 3 Chapter 4 Chapter 5 Chapter 6 Chapter 7 Chapter 8 Chapter 9 Thiol-disulfide oxidoreductase modules in the low-gc Grampositive bacteria Overflow of a hyper-produced secretory protein from the Bacillus Sec pathway into the Tat pathway for protein secretion as revealed by proteogenomics Contributions of the pre- and pro-regions of a Staphylococcus hyicus lipase to secretion of a heterologous protein by Bacillus subtilis Modulation of thiol-disulfide oxidoreductases for increased production of disulfide bond-containing proteins in Bacillus Thioredoxin A active site mutants form mixed disulfide dimers that resemble enzyme-substrate reaction intermediates MscL of Bacillus subtilis prevents selective release of cytoplasmic proteins in a hypotonic environment Immunity to the bacteriocin sublancin 168 is determined by the SunI (YolF) protein of Bacillus subtilis The large mechano-sensitive channel (MscL) determines bacterial sensitivity to the bacteriocin sublancin Chapter 10 Summary and general discussion 191 Chapter 11 Reference list 203 Appendices I. Nederlandse samenvatting voor de leek (Summary in Dutch) II. Dankwoord (Acknowledgements) III. List of publications

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