Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production
Abstract Background Mitochondrial carriers (MCs) can deeply affect the intracellular flux distribution of metabolic pathways. The manipulation of their expression level, to redirect the flux toward the production of a molecule of interest, is an attractive target for the metabolic engineering of euk...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-07-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-023-02137-8 |
| _version_ | 1797778664847310848 |
|---|---|
| author | Eugenia Messina Camilla Pires de Souza Claudia Cappella Simona Nicole Barile Pasquale Scarcia Isabella Pisano Luigi Palmieri Jean-Marc Nicaud Gennaro Agrimi |
| author_facet | Eugenia Messina Camilla Pires de Souza Claudia Cappella Simona Nicole Barile Pasquale Scarcia Isabella Pisano Luigi Palmieri Jean-Marc Nicaud Gennaro Agrimi |
| author_sort | Eugenia Messina |
| collection | DOAJ |
| description | Abstract Background Mitochondrial carriers (MCs) can deeply affect the intracellular flux distribution of metabolic pathways. The manipulation of their expression level, to redirect the flux toward the production of a molecule of interest, is an attractive target for the metabolic engineering of eukaryotic microorganisms. The non-conventional yeast Yarrowia lipolytica is able to use a wide range of substrates. As oleaginous yeast, it directs most of the acetyl-CoA therefrom generated towards the synthesis of lipids, which occurs in the cytoplasm. Among them, the odd-chain fatty acids (OCFAs) are promising microbial-based compounds with several applications in the medical, cosmetic, chemical and agricultural industries. Results In this study, we have identified the MC involved in the Carnitine/Acetyl-Carnitine shuttle in Y. lipolytica, YlCrc1. The Y. lipolytica Ylcrc1 knock-out strain failed to grow on ethanol, acetate and oleic acid, demonstrating the fundamental role of this MC in the transport of acetyl-CoA from peroxisomes and cytoplasm into mitochondria. A metabolic engineering strategy involving the deletion of YlCRC1, and the recombinant expression of propionyl-CoA transferase from Ralstonia eutropha (RePCT), improved propionate utilization and its conversion into OCFAs. These genetic modifications and a lipogenic medium supplemented with glucose and propionate as the sole carbon sources, led to enhanced accumulation of OCFAs in Y. lipolytica. Conclusions The Carnitine/Acetyl-Carnitine shuttle of Y. lipolytica involving YlCrc1, is the sole pathway for transporting peroxisomal or cytosolic acetyl-CoA to mitochondria. Manipulation of this carrier can be a promising target for metabolic engineering approaches involving cytosolic acetyl-CoA, as demonstrated by the effect of YlCRC1 deletion on OCFAs synthesis. |
| first_indexed | 2024-03-12T23:19:48Z |
| format | Article |
| id | doaj.art-ddbe986bdb884c52a1d41ac8ac7b4a04 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-03-12T23:19:48Z |
| publishDate | 2023-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-ddbe986bdb884c52a1d41ac8ac7b4a042023-07-16T11:32:05ZengBMCMicrobial Cell Factories1475-28592023-07-0122111610.1186/s12934-023-02137-8Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid productionEugenia Messina0Camilla Pires de Souza1Claudia Cappella2Simona Nicole Barile3Pasquale Scarcia4Isabella Pisano5Luigi Palmieri6Jean-Marc Nicaud7Gennaro Agrimi8Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”Université Paris-Saclay, INRAE, Micalis InstituteDepartment of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”Université Paris-Saclay, INRAE, Micalis InstituteDepartment of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”Abstract Background Mitochondrial carriers (MCs) can deeply affect the intracellular flux distribution of metabolic pathways. The manipulation of their expression level, to redirect the flux toward the production of a molecule of interest, is an attractive target for the metabolic engineering of eukaryotic microorganisms. The non-conventional yeast Yarrowia lipolytica is able to use a wide range of substrates. As oleaginous yeast, it directs most of the acetyl-CoA therefrom generated towards the synthesis of lipids, which occurs in the cytoplasm. Among them, the odd-chain fatty acids (OCFAs) are promising microbial-based compounds with several applications in the medical, cosmetic, chemical and agricultural industries. Results In this study, we have identified the MC involved in the Carnitine/Acetyl-Carnitine shuttle in Y. lipolytica, YlCrc1. The Y. lipolytica Ylcrc1 knock-out strain failed to grow on ethanol, acetate and oleic acid, demonstrating the fundamental role of this MC in the transport of acetyl-CoA from peroxisomes and cytoplasm into mitochondria. A metabolic engineering strategy involving the deletion of YlCRC1, and the recombinant expression of propionyl-CoA transferase from Ralstonia eutropha (RePCT), improved propionate utilization and its conversion into OCFAs. These genetic modifications and a lipogenic medium supplemented with glucose and propionate as the sole carbon sources, led to enhanced accumulation of OCFAs in Y. lipolytica. Conclusions The Carnitine/Acetyl-Carnitine shuttle of Y. lipolytica involving YlCrc1, is the sole pathway for transporting peroxisomal or cytosolic acetyl-CoA to mitochondria. Manipulation of this carrier can be a promising target for metabolic engineering approaches involving cytosolic acetyl-CoA, as demonstrated by the effect of YlCRC1 deletion on OCFAs synthesis.https://doi.org/10.1186/s12934-023-02137-8Yarrowia lipolyticaMitochondrial carrierCarnitine/Acetyl-Carnitine shuttleAcetyl-CoAMetabolic engineeringOdd-chain fatty acids (OCFAs) |
| spellingShingle | Eugenia Messina Camilla Pires de Souza Claudia Cappella Simona Nicole Barile Pasquale Scarcia Isabella Pisano Luigi Palmieri Jean-Marc Nicaud Gennaro Agrimi Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production Microbial Cell Factories Yarrowia lipolytica Mitochondrial carrier Carnitine/Acetyl-Carnitine shuttle Acetyl-CoA Metabolic engineering Odd-chain fatty acids (OCFAs) |
| title | Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production |
| title_full | Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production |
| title_fullStr | Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production |
| title_full_unstemmed | Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production |
| title_short | Genetic inactivation of the Carnitine/Acetyl-Carnitine mitochondrial carrier of Yarrowia lipolytica leads to enhanced odd-chain fatty acid production |
| title_sort | genetic inactivation of the carnitine acetyl carnitine mitochondrial carrier of yarrowia lipolytica leads to enhanced odd chain fatty acid production |
| topic | Yarrowia lipolytica Mitochondrial carrier Carnitine/Acetyl-Carnitine shuttle Acetyl-CoA Metabolic engineering Odd-chain fatty acids (OCFAs) |
| url | https://doi.org/10.1186/s12934-023-02137-8 |
| work_keys_str_mv | AT eugeniamessina geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT camillapiresdesouza geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT claudiacappella geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT simonanicolebarile geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT pasqualescarcia geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT isabellapisano geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT luigipalmieri geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT jeanmarcnicaud geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction AT gennaroagrimi geneticinactivationofthecarnitineacetylcarnitinemitochondrialcarrierofyarrowialipolyticaleadstoenhancedoddchainfattyacidproduction |