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

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

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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.
Original languageEnglish
Title of host publicationHydrocarbon and Lipid Microbiology Protocols : Synthetic and Systems Biology - Applications
PublisherSpringer
Publication date2015
Pages23-52
ISBN (Print)978-3-662-49126-3
ISBN (Electronic)978-3-662-49127-0
DOIs
Publication statusPublished - 2015
Externally publishedYes
SeriesSpringer Protocols Handbooks
ISSN1949-2448

Cite this

Vickers, C. E., Bongers, M., F. Bydder, S., Chrysanthopoulos, P., & Hodson, M. P. (2015). Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast. In Hydrocarbon and Lipid Microbiology Protocols: Synthetic and Systems Biology - Applications (pp. 23-52). Springer. Springer Protocols Handbooks https://doi.org/10.1007/8623_2015_107
Vickers, Claudia E. ; Bongers, Mareike ; F. Bydder, Sarah ; Chrysanthopoulos, Panagiotis ; Hodson, Mark P. / Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast. Hydrocarbon and Lipid Microbiology Protocols: Synthetic and Systems Biology - Applications. Springer, 2015. pp. 23-52 (Springer Protocols Handbooks).
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Vickers, CE, Bongers, M, F. Bydder, S, Chrysanthopoulos, P & Hodson, MP 2015, Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast. in Hydrocarbon and Lipid Microbiology Protocols: Synthetic and Systems Biology - Applications. Springer, Springer Protocols Handbooks, pp. 23-52. https://doi.org/10.1007/8623_2015_107

Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast. / Vickers, Claudia E.; Bongers, Mareike; F. Bydder, Sarah; Chrysanthopoulos, Panagiotis; Hodson, Mark P.

Hydrocarbon and Lipid Microbiology Protocols: Synthetic and Systems Biology - Applications. Springer, 2015. p. 23-52 (Springer Protocols Handbooks).

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

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AU - Hodson, Mark P.

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AB - 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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PB - Springer

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Vickers CE, Bongers M, F. Bydder S, Chrysanthopoulos P, Hodson MP. Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast. In Hydrocarbon and Lipid Microbiology Protocols: Synthetic and Systems Biology - Applications. Springer. 2015. p. 23-52. (Springer Protocols Handbooks). https://doi.org/10.1007/8623_2015_107