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10completethesis - PSEUDOMONAS AS A MICROBIAL ENZYME...

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P SEUDOMONAS AS A MICROBIAL ENZYME FACTORY The source of industrially potent enzymes and the host for heterologous enzyme production Joanna Krzeslak 2009
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R IJKSUNIVERSITEIT G RONINGEN P SEUDOMONAS AS A MICROBIAL ENZYME FACTORY The source of industrially potent enzymes and the host for heterologous enzyme production 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 17 april 2009 om 14.45 uur door Joanna Kamila Krzeslak geboren op 17 augustus 1978 te Olsztyn, Polen
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P ROMOTOR : Prof. dr. W.J. Quax B EOORDELINGSCOMMISSIE : Prof. dr. A.A.M Filloux Prof. dr. K-E. Jaeger Prof. dr. O. Kayser
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The life which is unexamined is not worth living. Plato The shoe that fits one person pinches another; there is no recipe for living that suits all cases. Carl Jung Ukochanym
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P ARANIMFEN : Joanna Majchrzykiewicz Iryna Monastyrska The studies described in this thesis were performed at the Department of Pharmaceutical Biology of the University of Groningen and were partly funded by the EU grant QLK3-CT-2002- 02086. Printed by Ridderprint B. V., Ridderkerk. ISBN printed version: 978-90-367-3783-8 ISBN digital version: 978-90-367-3784-5
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C C ONTENTS C HAPTER 1 Introduction and scope of the thesis 9 C HAPTER 2 Quorum quenching acylases in Pseudomonas aeruginosa 27 C HAPTER 3 Lipase expression in Pseudomonas alcaligenes is under the control of a two-component regulatory system 47 C HAPTER 4 LipR, a response regulator, directly controls the lipase gene expression in Pseudomonas alcaligenes 69 C HAPTER 5 Heterologous production of Escherichia coli penicillin G acylase in Pseudomonas aeruginosa 85 C HAPTER 6 Cloning, sequence analysis, and expression of PA1893, a putative acylase, from Pseudomonas aeruginosa PAO1 105 C HAPTER 7 Summary, general discussion and future perspectives 129 Nederlandse samenvatting 137 Acknowledgements 145
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C C HAPTER 1 1 Introduction and scope of the thesis
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General introduction 11 Introduction and scope of the thesis Organization of life The tree of life is made up of three distinct domains: eukaryotes, bacteria, and archaea 1,2 . The eukaryotes represent organisms whose cells are organized into complex structures enclosed within membranes. The defining membrane-bound structure that distinguishes them from other forms of life is the nucleus containing most of the cell’s genetic material. Eukaryotes include animals, plants, fungi and a rich variety of microorganisms known as protists. On the contrary, archaea and bacteria are prokaryotes and have no cell nucleus or any other organelles within their cells. Archaea are an ancient form of life generally quite similar in size and shape to bacteria. They can be differentiated from bacteria based on the sequences of ribosomal RNA (rRNA) genes 3 . The bacteria can be classified as Gram-positive or Gram- negative bacteria based on differences in their cell wall. Gram-positive bacteria possess a thick cell wall containing many layers of peptidoglycan and teichoic acid. In contrast, Gram-negative
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