Details
Original language | English |
---|---|
Pages (from-to) | 4038-4046 |
Number of pages | 9 |
Journal | CHEMCATCHEM |
Volume | 13 |
Issue number | 18 |
Early online date | 12 Jul 2021 |
Publication status | Published - 17 Sept 2021 |
Externally published | Yes |
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
- Chemical Engineering(all)
- Catalysis
- Chemistry(all)
- Physical and Theoretical Chemistry
- Chemistry(all)
- Organic Chemistry
- Chemistry(all)
- Inorganic Chemistry
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In: CHEMCATCHEM, Vol. 13, No. 18, 17.09.2021, p. 4038-4046.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Optimization and Engineering of Fatty Acid Photodecarboxylase for Substrate Specificity
AU - Santner, Paul
AU - Szabó, László Krisztián
AU - Chanquia, Santiago Nahuel
AU - Merrild, Aske Høj
AU - Hollmann, Frank
AU - Kara, Selin
AU - Eser, Bekir Engin
N1 - Funding Information: The authors thank the Novo Nordisk Foundation, Light‐BioFuels project, grant No. NNF19OC0057522. Publisher Copyright: © 2021 Wiley-VCH GmbH
PY - 2021/9/17
Y1 - 2021/9/17
N2 - 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.
AB - 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.
KW - Biocatalysis
KW - Drop-in Biofuels
KW - Fatty Acid Photodecarboxylase
KW - Photoenzyme
KW - Protein Engineering
UR - http://www.scopus.com/inward/record.url?scp=85111354890&partnerID=8YFLogxK
U2 - 10.1002/cctc.202100840
DO - 10.1002/cctc.202100840
M3 - Article
AN - SCOPUS:85111354890
VL - 13
SP - 4038
EP - 4046
JO - CHEMCATCHEM
JF - CHEMCATCHEM
SN - 1867-3880
IS - 18
ER -