Biocompatible fluorocarbon liquid underlays for in situ extraction of isoprenoids from microbial cultures

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5 Scopus citations

Abstract

Microbial production of heterologous metabolites is now a mature technology in many host organisms, opening new avenues for green production processes of specialty chemicals. At lab scale, petroleum-based hydrophobic bio-compatible solvents like dodecane can be used as a second phase on top of microbial cultures to act as a physical sink for heterologous hydrocarbon products like isoprenoids. However, this approach has significant drawbacks at scale due to the difficulty of handling solvents and their potential contamination with unwanted byproducts of their manufacture. We discovered that synthetic perfluorocarbon liquids (FCs), commonly used for heat transfer, can also act as physical sinks for microbially produced isoprenoid compounds. FCs are stable, inert, and are amenable to direct liquid–liquid extraction with alcohols for rapid product isolation. These liquids are more dense than water and form a lower phase to microbial cultures rather than an upper phase as with other solvents. Their ability to form an under-layer or ‘underlay’ also enables the cultivation of microbes directly at the FC–culture medium interface via gravity settling, which could open their application for filamentous or mat-forming organisms. We present comparisons of the isoprenoid extraction potential of three commercial FCs: FC-3283, FC-40, and FC-770 with engineered green microalga cultures producing patchoulol, taxadiene, casbene, or 13R(+) manoyl oxide. We demonstrate that FCs are promising alternatives to traditional solvents and open new avenues in bio-process design for microbial heterologous metabolite milking.
Original languageEnglish (US)
Pages (from-to)16632-16639
Number of pages8
JournalRSC ADVANCES
Volume12
Issue number26
DOIs
StatePublished - Jun 6 2022

Bibliographical note

KAUST Repository Item: Exported on 2022-06-15
Acknowledged KAUST grant number(s): 4238
Acknowledgements: We would like to express special thanks to Najeh Kharbatia of the KAUST Analytical Core Labs for helpful early discussions, Chandrasekaren Lakshmipathy, Abdulkhalik Khalifa, and Abdullah Alabdullatif of KAUST Lab Equipment Maintenance (LEM) team for assistance in upgrading and initializing the GC-FID-MS unit. The authors acknowledge Prof. Dr Ralph Bock and Dr Juliane Neupert for providing C. reinhardtii UVM4, obtained under material transfer agreement between KAUST and the Max-Planck-Institut für Molekulare Pflanzenphysiologie Potsdam. Diterpenoid production strains of C. reinhardtii used in this work were developed by KJL in the lab of Prof. Dr Olaf Kruse at the Center for Biotechnology, Universität Bielefeld as referenced in the manuscript. The research reported in this publication was supported by the KAUST Impact Acceleration Funds program (grant 4238) and KAUST baseline funding awarded to Kyle Lauersen.

ASJC Scopus subject areas

  • General Chemical Engineering
  • General Chemistry

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