Optimization and Engineering of Fatty Acid Photodecarboxylase for Substrate Specificity

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Paul Santner
  • László Krisztián Szabó
  • Santiago Nahuel Chanquia
  • Aske Høj Merrild
  • Frank Hollmann
  • Selin Kara
  • Bekir Engin Eser

External Research Organisations

  • Aarhus University
  • Delft University of Technology
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Details

Original languageEnglish
Pages (from-to)4038-4046
Number of pages9
JournalCHEMCATCHEM
Volume13
Issue number18
Early online date12 Jul 2021
Publication statusPublished - 17 Sept 2021
Externally publishedYes

Abstract

Fatty acid photodecarboxylase (FAP) is one of the few photoenzymes in nature. The ability of FAP to convert fatty acids into alka(e)nes without the need for reducing equivalents put this enzyme into spotlight for biocatalytic applications. Although it has been discovered only a few years ago, many studies already emerged demonstrating its potential in areas from biofuel production and enzymatic kinetic resolution to being a critical component of multi-enzyme cascades. While there have been few protein engineering studies for modulating activity of FAP towards very short chain fatty acids, no study has yet addressed substrate selectivity within the medium to long chain fatty acid range, where FAP shows great promise for the synthesis of drop-in biofuels from ubiquitous fatty acids with chain lengths from C12 to C18. Here, after determining optimum expression and assay conditions for FAP, we screened 22 rationally designed mutant enzymes towards four naturally abundant fatty acid substrates; C12 : 0, C16 : 0, C18 : 0 and C18 : 1. Depending on the type of the exchanged amino acid, we observed selectivity shifts towards shorter or longer chains, compared to wild type enzyme. Notably, we obtained two groups of mutants; one group with high selectivity towards only C18 : 0, and another group that is selective towards C12 : 0 substrate. Moreover, we measured light and thermal stability of the wild type enzyme as well as the light stability of a mutant engineered for selectivity.

Keywords

    Biocatalysis, Drop-in Biofuels, Fatty Acid Photodecarboxylase, Photoenzyme, Protein Engineering

ASJC Scopus subject areas

Cite this

Optimization and Engineering of Fatty Acid Photodecarboxylase for Substrate Specificity. / Santner, Paul; Szabó, László Krisztián; Chanquia, Santiago Nahuel et al.
In: CHEMCATCHEM, Vol. 13, No. 18, 17.09.2021, p. 4038-4046.

Research output: Contribution to journalArticleResearchpeer review

Santner, P, Szabó, LK, Chanquia, SN, Merrild, AH, Hollmann, F, Kara, S & Eser, BE 2021, 'Optimization and Engineering of Fatty Acid Photodecarboxylase for Substrate Specificity', CHEMCATCHEM, vol. 13, no. 18, pp. 4038-4046. https://doi.org/10.1002/cctc.202100840
Santner, P., Szabó, L. K., Chanquia, S. N., Merrild, A. H., Hollmann, F., Kara, S., & Eser, B. E. (2021). Optimization and Engineering of Fatty Acid Photodecarboxylase for Substrate Specificity. CHEMCATCHEM, 13(18), 4038-4046. https://doi.org/10.1002/cctc.202100840
Santner P, Szabó LK, Chanquia SN, Merrild AH, Hollmann F, Kara S et al. Optimization and Engineering of Fatty Acid Photodecarboxylase for Substrate Specificity. CHEMCATCHEM. 2021 Sept 17;13(18):4038-4046. Epub 2021 Jul 12. doi: 10.1002/cctc.202100840
Santner, Paul ; Szabó, László Krisztián ; Chanquia, Santiago Nahuel et al. / Optimization and Engineering of Fatty Acid Photodecarboxylase for Substrate Specificity. In: CHEMCATCHEM. 2021 ; Vol. 13, No. 18. pp. 4038-4046.
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abstract = "Fatty acid photodecarboxylase (FAP) is one of the few photoenzymes in nature. The ability of FAP to convert fatty acids into alka(e)nes without the need for reducing equivalents put this enzyme into spotlight for biocatalytic applications. Although it has been discovered only a few years ago, many studies already emerged demonstrating its potential in areas from biofuel production and enzymatic kinetic resolution to being a critical component of multi-enzyme cascades. While there have been few protein engineering studies for modulating activity of FAP towards very short chain fatty acids, no study has yet addressed substrate selectivity within the medium to long chain fatty acid range, where FAP shows great promise for the synthesis of drop-in biofuels from ubiquitous fatty acids with chain lengths from C12 to C18. Here, after determining optimum expression and assay conditions for FAP, we screened 22 rationally designed mutant enzymes towards four naturally abundant fatty acid substrates; C12 : 0, C16 : 0, C18 : 0 and C18 : 1. Depending on the type of the exchanged amino acid, we observed selectivity shifts towards shorter or longer chains, compared to wild type enzyme. Notably, we obtained two groups of mutants; one group with high selectivity towards only C18 : 0, and another group that is selective towards C12 : 0 substrate. Moreover, we measured light and thermal stability of the wild type enzyme as well as the light stability of a mutant engineered for selectivity.",
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AU - Szabó, László Krisztián

AU - Chanquia, Santiago Nahuel

AU - Merrild, Aske Høj

AU - Hollmann, Frank

AU - Kara, Selin

AU - Eser, Bekir Engin

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