Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA
Malonyl-CoA is an energy-rich molecule formed by the ATP-dependent carboxylation of acetyl coenzyme A catalyzed by acetyl-CoA carboxylase. This molecule is an important precursor for many biotechnologically interesting compounds such as flavonoids, polyketides, and fatty acids. The yeast Saccharomyc...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-01-01
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| Series: | Computational and Structural Biotechnology Journal |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037022000277 |
| _version_ | 1797978228093091840 |
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| author | Humberto Pereira Flávio Azevedo Lucília Domingues Björn Johansson |
| author_facet | Humberto Pereira Flávio Azevedo Lucília Domingues Björn Johansson |
| author_sort | Humberto Pereira |
| collection | DOAJ |
| description | Malonyl-CoA is an energy-rich molecule formed by the ATP-dependent carboxylation of acetyl coenzyme A catalyzed by acetyl-CoA carboxylase. This molecule is an important precursor for many biotechnologically interesting compounds such as flavonoids, polyketides, and fatty acids. The yeast Saccharomyces cerevisiae remains one of the preferred cell factories, but has a limited capacity to produce malonyl-CoA compared to oleaginous organisms. We developed a new S. cerevisiae strain with a conditional allele of ACC1, the essential acetyl-CoA carboxylase (ACC) gene, as a tool to test heterologous genes for complementation. Yarrowia lipolytica is an oleaginous yeast with a higher capacity for lipid production than S. cerevisiae, possibly due to a higher capacity to produce malonyl-CoA. Measuring relative intracellular malonyl-CoA levels with an in-vivo biosensor confirmed that expression of Y. lipolytica ACC in S. cerevisiae leads to a higher accumulation of malonyl-CoA compared with overexpression of the native gene from an otherwise identical vector. The higher accumulation was generally accompanied by a decreased growth rate. Concomitant expression of both the homologous and heterologous ACC1 genes eliminated the growth defect, with a marginal reduction of malonyl-CoA accumulation. |
| first_indexed | 2024-04-11T05:20:29Z |
| format | Article |
| id | doaj.art-8a978b636ba84e70b4466555d33592d5 |
| institution | Directory Open Access Journal |
| issn | 2001-0370 |
| language | English |
| last_indexed | 2024-04-11T05:20:29Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Computational and Structural Biotechnology Journal |
| spelling | doaj.art-8a978b636ba84e70b4466555d33592d52022-12-24T04:51:19ZengElsevierComputational and Structural Biotechnology Journal2001-03702022-01-0120779787Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoAHumberto Pereira0Flávio Azevedo1Lucília Domingues2Björn Johansson3CBMA - Center of Molecular and Environmental Biology EngineeringCBMA - Center of Molecular and Environmental Biology EngineeringCEB - Centre of Biological Engineering, University of Minho, Campus de Gualtar, Braga, 4710-057, PortugalCBMA - Center of Molecular and Environmental Biology Engineering; Corresponding author.Malonyl-CoA is an energy-rich molecule formed by the ATP-dependent carboxylation of acetyl coenzyme A catalyzed by acetyl-CoA carboxylase. This molecule is an important precursor for many biotechnologically interesting compounds such as flavonoids, polyketides, and fatty acids. The yeast Saccharomyces cerevisiae remains one of the preferred cell factories, but has a limited capacity to produce malonyl-CoA compared to oleaginous organisms. We developed a new S. cerevisiae strain with a conditional allele of ACC1, the essential acetyl-CoA carboxylase (ACC) gene, as a tool to test heterologous genes for complementation. Yarrowia lipolytica is an oleaginous yeast with a higher capacity for lipid production than S. cerevisiae, possibly due to a higher capacity to produce malonyl-CoA. Measuring relative intracellular malonyl-CoA levels with an in-vivo biosensor confirmed that expression of Y. lipolytica ACC in S. cerevisiae leads to a higher accumulation of malonyl-CoA compared with overexpression of the native gene from an otherwise identical vector. The higher accumulation was generally accompanied by a decreased growth rate. Concomitant expression of both the homologous and heterologous ACC1 genes eliminated the growth defect, with a marginal reduction of malonyl-CoA accumulation.http://www.sciencedirect.com/science/article/pii/S2001037022000277Saccharomyces cerevisiaeACC1Fatty acidsMalonyl-CoABiosensorYarrowia lipolytica |
| spellingShingle | Humberto Pereira Flávio Azevedo Lucília Domingues Björn Johansson Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA Computational and Structural Biotechnology Journal Saccharomyces cerevisiae ACC1 Fatty acids Malonyl-CoA Biosensor Yarrowia lipolytica |
| title | Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA |
| title_full | Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA |
| title_fullStr | Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA |
| title_full_unstemmed | Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA |
| title_short | Expression of Yarrowia lipolytica acetyl-CoA carboxylase in Saccharomyces cerevisiae and its effect on in-vivo accumulation of Malonyl-CoA |
| title_sort | expression of yarrowia lipolytica acetyl coa carboxylase in saccharomyces cerevisiae and its effect on in vivo accumulation of malonyl coa |
| topic | Saccharomyces cerevisiae ACC1 Fatty acids Malonyl-CoA Biosensor Yarrowia lipolytica |
| url | http://www.sciencedirect.com/science/article/pii/S2001037022000277 |
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