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Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast

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Hydrocarbon and Lipid Microbiology Protocols

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

Isoprenoids (a.k.a. terpenes/terpenoids) are the largest group of natural products. They fulfil a wide variety of both essential and non-essential roles in biology; many isoprenoids also have useful industrial applications. In recent years, there has been a significant focus on metabolic engineering of various isoprenoids in microbial cells. Here, we describe methods for lab-scale culturing, sampling and analytics of different isoprenoid classes using specific examples from the following classes: (1) highly volatile isoprenoids that sequester into culture headspaces, e.g. the hemiterpene isoprene; (2) volatile isoprenoids that can be collected in a non-toxic organic phase, e.g. the monoterpene limonene; and (3) non-volatile isoprenoids that accumulate in the cell, e.g. the carotenoid lycopene. Production methods are provided for isoprene, limonene, and lycopene as examples for each class. Specific analytical methods are provided for isoprene, limonene, terpinolene, caryophyllene, amorphadiene, linalool, nerolidol, pinene and lycopene. We focus on yeast and Escherichia coli as production organisms. The protocols can be modified for other organisms and products as appropriate.

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Abbreviations

CDM:

Chemically defined medium

ECMet:

Extracellular metabolites

GC–MS:

Gas chromatography–mass spectrometry

HPLC:

High pressure/performance liquid chromatography

ISTD:

Internal standard

LLOQ:

Lower limit of quantitation

LOD:

Limit of detection

OD:

Optical density

PTFE:

Polytetrafluoroethylene (commonly identified as Teflon®, a DuPont brand name)

rpm:

revolutions per minute

SIM:

Selected ion monitoring

TB:

Terrific broth

TIC:

Total ion current (chromatogram)

ULOQ:

Upper limit of quantitation

YPD:

Yeast extract peptone dextrose (a.k.a. YEPD)

YPDG:

Yeast extract peptone dextrose galactose

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Author information

Authors and Affiliations

  1. Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, 4072, Australia

    Claudia E. Vickers, Mareike Bongers, Sarah F. Bydder, Panagiotis Chrysanthopoulos & Mark P. Hodson

  2. Metabolomics Australia Queensland Node, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, 4072, Australia

    Panagiotis Chrysanthopoulos & Mark P. Hodson

Authors
  1. Claudia E. Vickers
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  2. Mareike Bongers
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  3. Sarah F. Bydder
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  4. Panagiotis Chrysanthopoulos
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  5. Mark P. Hodson
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Corresponding author

Correspondence to Claudia E. Vickers .

Editor information

Editors and Affiliations

  1. Dept. Biological Sciences, University of Essex, Colchester, Essex, United Kingdom

    Terry J. McGenity

  2. Braunschweig, Germany

    Kenneth N. Timmis

  3. Dept. Biologia, Univ de les Illes Balears, Palma de Mallorca, Spain

    Balbina Nogales Fernández

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© 2015 Springer-Verlag Berlin Heidelberg

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Vickers, C.E., Bongers, M., Bydder, S.F., Chrysanthopoulos, P., Hodson, M.P. (2015). Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast. In: McGenity, T., Timmis, K., Nogales Fernández, B. (eds) Hydrocarbon and Lipid Microbiology Protocols. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8623_2015_107

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  • DOI: https://doi.org/10.1007/8623_2015_107

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  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-49126-3

  • Online ISBN: 978-3-662-49127-0

  • eBook Packages: Springer Protocols

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