University of Groningen Selection of novel lipases and esterases for enantioselective biocatalysis Dröge, Melloney Joyce 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: 2004 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Dröge, M. J. (2004). Selection of novel lipases and esterases for enantioselective biocatalysis 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): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date: 04-12-2017
Selection of novel lipases and esterases for enantioselective biocatalysis
Rijksuniversiteit Groningen Selection of novel lipases and esterases for enantioselective biocatalysis Proefschrift ter verkrijging van het doctoraat in de Wiskunde en Natuurwetenschappen aan de Rijksuniversiteit Groningen op gezag van de Rector Magnificus, dr. F. Zwarts, in het openbaar te verdedigen op vrijdag 13 februari 2004 om 16.15 uur door Melloney Joyce Dröge geboren op 22 juli 1975 te Groningen
Onze drie-dimensionale ruimte is de enige realiteit die wij kennen. Het twee-dimensionale is even fictief als het vier-dimensionale, want niets is vlak M. C. Escher Aan mijn ouders Voor Folgert
Promotor: Prof. dr. W. J. Quax Beoordelingscommissie: Prof. dr. B. W. Dijkstra Prof. dr. K-E. Jaeger Prof. dr. J. Zaagsma ISBN-nummer: 90-901-7661-6
Paranimfen: Marc Dröge Ykelien Boersma This research was performed at the Department of Pharmaceutical Biology (University of Groningen, The Netherlands), and was funded by the European Commission under proposal number BIO4-98-0249 and QLK3-CT-2001-00519. Financial support by the Dr. Ir. Van de Laar Stichting, MSD, and the Groningen University Institute for Drug Exploration (GUIDE) for the printing of this thesis is gratefully acknowledged. Printing: Ridderprint BV (Ridderkerk, The Netherlands). 2004 M. J. Dröge. All right reserved. No part of this book may be reproduced or transmitted in any forms or by any means, without permission from the author. Cover: M. C. Escher s Drawing Hands 2003 Cordon Art, Baarn, Holland. All rights reserved.
Contents Chapter 1: Introduction...9 Chapter 2: Paralogous gene analysis reveals a highly enantioselective 1,2- O-isopropylideneglycerol caprylate esterase of Bacillus subtilis...27 Chapter 3: Chapter 4: Chapter 5: Chapter 6: Chapter 7: Comparison and functional characterisation of three homologous intracellular carboxylesterases of Bacillus subtilis...41 Chiral gas chromatography for the determination of 1,2-O-isopropylidene-sn-glycerol stereoisomers...61 Phage display of an intracellular carboxylesterase of Bacillus subtilis: a comparison of the Sec and Tat pathway capabilities...73 Binding of phage displayed Bacillus subtilis lipase A to a phosphonate suicide inhibitor...91 Immobilisation of chiral enzyme inhibitors on solid supports by amide-forming coupling and olefin metathesis...105 Chapter 8: Directed evolution of Bacillus subtilis lipase A using enantiomeric phosphonate inhibitors: crystal structures and phage display selection...123 Chapter 9: Summary, general discussion and perspectives...143 References... 153 Nederlandse samenvatting...167 List of publications...171 Dankwoord...173