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Bacillus subtilis spore surface display of photodecarboxylase for the transformation of lipids to hydrocarbons

Marianna Karava, Peter Gockel, Johannes Kabisch
doi: https://doi.org/10.1101/2020.08.30.273821
Marianna Karava
1Computer-aided Synthetic Biology, Institute for Biology, Technische Universität Darmstadt, Darmstadt, Germany
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Peter Gockel
1Computer-aided Synthetic Biology, Institute for Biology, Technische Universität Darmstadt, Darmstadt, Germany
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Johannes Kabisch
1Computer-aided Synthetic Biology, Institute for Biology, Technische Universität Darmstadt, Darmstadt, Germany
2Centre for Synthetic Biology, Technische Universität Darmstadt, Darmstadt, Germany
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  • For correspondence: johannes@kabisch-lab.de
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Abstract

The display of enzymes on the surface of spores allows the rapid and very simple biotechnological production of immobilized enzymes. Here we describe the development of a Bacillus subtilis spore display platform and its application to produce hydrocarbons from lipids obtained from the oleaginous yeasts Yarrowia lipolytica, Cutaneotrichosporon oleaginosus as well as olive oil.

Lipid hydrolysis was examined in a bienzymatic one-pot cascade using a commercially immobilized lipase (RO lipase) as well as spores with and without additional heterologous lipase expression. Decarboxylation of the released fatty acids was achieved displaying a photodecarboxylase (CvFAP) on the spore surface. Differences in composition of the formed hydrocarbons were observed depending on the lipids source. Using 3D printed lighting equipment titers of up to 64.0 ± 5.6 mg/L hydrocarbons were produced.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://gitlab.com/kabischlab.de/led-labware-cvfap

  • Abbreviations
    SDW
    spore dry weight
    TAGs
    triacylglycerides
    HC
    hydrocarbon
    Co. oil
    natural oil produced by Cutaneotrichosporon oleaginosus
    Yali. oil
    natural oil produced by Yarrowia lipolytica
    RO
    Rhizopus oryzae
    WO
    without
    SG
    Schaffers glucose
  • Copyright 
    The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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    Bacillus subtilis spore surface display of photodecarboxylase for the transformation of lipids to hydrocarbons
    Marianna Karava, Peter Gockel, Johannes Kabisch
    bioRxiv 2020.08.30.273821; doi: https://doi.org/10.1101/2020.08.30.273821
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    Bacillus subtilis spore surface display of photodecarboxylase for the transformation of lipids to hydrocarbons
    Marianna Karava, Peter Gockel, Johannes Kabisch
    bioRxiv 2020.08.30.273821; doi: https://doi.org/10.1101/2020.08.30.273821

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