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University of Groningen Developing Bacillus subtilis as a versatile bioproduct platform for agricultural and pharmaceutical applications Song, Yafeng

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University of Groningen

Developing Bacillus subtilis as a versatile bioproduct platform for agricultural and

pharmaceutical applications

Song, Yafeng

DOI:

10.33612/diss.168189909

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: 2021

Link to publication in University of Groningen/UMCG research database

Citation for published version (APA):

Song, Y. (2021). Developing Bacillus subtilis as a versatile bioproduct platform for agricultural and pharmaceutical applications. University of Groningen. https://doi.org/10.33612/diss.168189909

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Developing Bacillus subtilis as a versatile

bioproduct platform for agricultural and

pharmaceutical applications

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The research described in this thesis was carried out in the Department of Chemical and Pharmaceutical Biology (Groningen Research Institute of Pharmacy, University of Groningen, The Netherlands) and was financially supported by the China Scholarship Council.

The research work was carried out according to the requirements of the Graduate School of Science, Faculty of Science and Engineering, University of Groningen, The Netherlands.

Printing of this thesis was financially supported by the University Library and the Graduate School of Science, Faculty of Science and Engineering, University of Groningen, The Netherlands.

Printing: Ridderprint BV, www.ridderprint.nl Cover design: Yafeng Song and Ridderprint

Copyright © 2021 Yafeng Song. All rights are reserved. No part of the thesis may be reproduced or transmitted in any form or by any means without the prior permission in writing of the author.

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Developing Bacillus subtilis as a

versatile bioproduct platform for

agricultural and pharmaceutical

applications

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 by the College of Deans.

This thesis will be defended in public on

Friday 7 May 2021 at 12.45 hours

by

Yafeng Song

born on 18 October 1990 in Hebei, China

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Supervisors

Prof. W.J. Quax

Prof. G.J. Poelarends

Assessment Committee

Prof. J. Stulke

Prof. J.M. van Dijl

Prof. F.J. Dekker

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CONTENTS

Chapter 1 Introduction and scope of this thesis

7

Chapter 2 High-efficiency secretion of β-mannanase in Bacillus subtilis through

protein synthesis and secretion optimization

23

Chapter 3

Positioning Bacillus subtilis as terpenoid cell factory

51

Chapter 4

Production of squalene in Bacillus subtilis by squalene synthases

screening and metabolic engineering

87

Chapter 5

Engineering of multiple modules to improve amorphadiene production

in Bacillus subtilis using CRISPR-Cas9

121

Chapter 6

Development and application of CRISPR based genetic tools in

Bacillus species

153

Chapter 7

Summary and future perspectives

185

Appendix

Acknowledgements

List of publications

About the author

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