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Pseudomonas Methods and Protocols

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Table of Contents

  1. Altmetric Badge
    Book Overview
  2. Altmetric Badge
    Chapter 1 Gene Transfer: Transduction
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    Chapter 2 Gene Transfer: Transformation/Electroporation
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    Chapter 3 Gene transfer: conjugation.
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    Chapter 4 Pseudomonas Bacteriophage Isolation and Production
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    Chapter 5 Genotyping Methods
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    Chapter 6 Drug Susceptibility Testing by Dilution Methods
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    Chapter 7 Plate-Based Assay for Swimming Motility in Pseudomonas aeruginosa
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    Chapter 8 Plate-Based Assay for Swarming Motility in Pseudomonas aeruginosa
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    Chapter 9 Motility Assay: Twitching Motility
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    Chapter 10 Qualitative and Quantitative Assays for Flagellum-Mediated Chemotaxis
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    Chapter 11 Microscopic Analysis: Morphotypes and Cellular Appendages
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    Chapter 12 Determination of Lipolytic Enzyme Activities
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    Chapter 13 Elastinolytic and Proteolytic Enzymes
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    Chapter 14 In vitro Assays to Monitor the Activity of Pseudomonas aeruginosa Type III Secreted Proteins
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    Chapter 15 Cell Fractionation
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    Chapter 16 Characterization of Molecular Interactions Using Isothermal Titration Calorimetry
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    Chapter 17 Proteomic analysis.
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    Chapter 18 Membrane Proteomics of Pseudomonas aeruginosa
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    Chapter 19 Construction of Pseudomonas aeruginosa Two-Hybrid Libraries for High-Throughput Assays
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    Chapter 20 Biosensors for Qualitative and Semiquantitative Analysis of Quorum Sensing Signal Molecules
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    Chapter 21 LC-MS/MS Quantitative Analysis of Quorum Sensing Signal Molecules
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    Chapter 22 LC/MS/MS-Based Quantitative Assay for the Secondary Messenger Molecule, c-di-GMP.
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    Chapter 23 Metabolic footprinting: extracellular metabolomic analysis.
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    Chapter 24 Pyoverdine and Pyochelin Measurements
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    Chapter 25 Measurement of Phenazines in Bacterial Cultures
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    Chapter 26 Extraction and Measurement of NAD(P) + and NAD(P)H
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    Chapter 27 Cyanide Measurements in Bacterial Culture and Sputum
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    Chapter 28 Monitoring iron uptake by siderophores.
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    Chapter 29 Exopolysaccharide quantification.
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    Chapter 30 Liquid Chromatography/Mass Spectrometry for the Identification and Quantification of Rhamnolipids
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    Chapter 31 LPS Quantitation Procedures
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    Chapter 32 Monitoring lectin interactions with carbohydrates.
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    Chapter 33 Mining the Pseudomonas Genome
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    Chapter 34 Identification of Bacterial Small RNAs by RNA Sequencing.
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    Chapter 35 Gene Amplification and qRT-PCR
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    Chapter 36 The Standard European Vector Architecture (SEVA) Plasmid Toolkit
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    Chapter 37 Chromosomal integration of transcriptional fusions.
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    Chapter 38 A Method to Capture Large DNA Fragments from Genomic DNA
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    Chapter 39 Transposon mutagenesis.
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    Chapter 40 Site-Directed Mutagenesis and Gene Deletion Using Reverse Genetics
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    Chapter 41 Signature-Tagged Mutagenesis
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    Chapter 42 Construction of a Pseudomonas aeruginosa Genomic DNA Library
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    Chapter 43 Pseudomonas Methods and Protocols
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    Chapter 44 Promoter fusions with optical outputs in individual cells and in populations.
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    Chapter 45 Chromatin Immunoprecipitation for ChIP-chip and ChIP-seq.
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    Chapter 46 Pseudomonas Methods and Protocols
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    Chapter 47 Methods for studying biofilm formation: flow cells and confocal laser scanning microscopy.
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    Chapter 48 Biofilm formation in the 96-well microtiter plate.
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    Chapter 49 Methods for Studying Biofilm Dispersal in Pseudomonas aeruginosa
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    Chapter 50 Pseudomonas aeruginosa PA14 Pathogenesis in Caenorhabditis elegans
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    Chapter 51 Assessing Pseudomonas aeruginosa Virulence Using a Nonmammalian Host: Dictyostelium discoideum
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    Chapter 52 Assessing Pseudomonas Virulence with Nonmammalian Host: Galleria mellonella
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    Chapter 53 Assessing Pseudomonas Virulence with the Nonmammalian Host Model: Arabidopsis thaliana
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    Chapter 54 Assessing Pseudomonas aeruginosa Persister/Antibiotic Tolerant Cells
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    Chapter 55 Assessing Pseudomonas Virulence with Nonmammalian Host: Zebrafish
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    Chapter 56 Assessing Pseudomonas Virulence with a Nonmammalian Host: Drosophila melanogaster.
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    Chapter 57 Assessing Pseudomonas Virulence Using Host Cells
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    Chapter 58 Assessing Pseudomonas aeruginosa Virulence and the Host Response Using Murine Models of Acute and Chronic Lung Infection
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    Chapter 59 Assessing Pseudomonas Virulence Using Mammalian Models: Acute Infection Model
  61. Altmetric Badge
    Chapter 60 Burn Mouse Models
Attention for Chapter 12: Determination of Lipolytic Enzyme Activities
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Chapter title
Determination of Lipolytic Enzyme Activities
Chapter number 12
Book title
Pseudomonas Methods and Protocols
Published in
Methods in molecular biology, January 2014
DOI 10.1007/978-1-4939-0473-0_12
Pubmed ID
Book ISBNs
978-1-4939-0472-3, 978-1-4939-0473-0

Karl-Erich Jaeger, Filip Kovacic, Jaeger, Karl-Erich, Kovacic, Filip


Pseudomonas aeruginosa is a versatile human opportunistic pathogen that produces and secretes an arsenal of enzymes, proteins and small molecules many of which serve as virulence factors. Notably, about 40 % of P. aeruginosa genes code for proteins of unknown function, among them more than 80 encoding putative, but still unknown lipolytic enzymes. This group of hydrolases (EC 3.1.1) is known already for decades, but only recently, several of these enzymes have attracted attention as potential virulence factors. Reliable and reproducible enzymatic activity assays are crucial to determine their physiological function and particularly assess their contribution to pathogenicity. As a consequence of the unique biochemical properties of lipids resulting in the formation of micellar structures in water, the reproducible preparation of substrate emulsions is strongly dependent on the method used. Furthermore, the physicochemical properties of the respective substrate emulsion may drastically affect the activities of the tested lipolytic enzymes. Here, we describe common methods for the activity determination of lipase, esterase, phospholipase, and lysophospholipase. These methods cover lipolytic activity assays carried out in vitro, with cell extracts or separated subcellular compartments and with purified enzymes. We have attempted to describe standardized protocols, allowing the determination and comparison of enzymatic activities of lipolytic enzymes from different sources. These methods should also encourage the Pseudomonas community to address the wealth of still unexplored lipolytic enzymes encoded and produced by P. aeruginosa.

Mendeley readers

The data shown below were compiled from readership statistics for 52 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Uruguay 1 2%
Unknown 51 98%

Demographic breakdown

Readers by professional status Count As %
Student > Master 13 25%
Student > Ph. D. Student 11 21%
Student > Bachelor 4 8%
Researcher 4 8%
Student > Doctoral Student 3 6%
Other 4 8%
Unknown 13 25%
Readers by discipline Count As %
Agricultural and Biological Sciences 10 19%
Biochemistry, Genetics and Molecular Biology 9 17%
Immunology and Microbiology 3 6%
Engineering 3 6%
Environmental Science 2 4%
Other 7 13%
Unknown 18 35%

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