Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3
Fatty acid desaturase enzymes play an essential role in the synthesis of unsaturated fatty acids. Pseudomonas sp. A3 was found to produce a large amount of palmitoleic and oleic acids after incubation at low temperatures. Using polymerase Chain Reaction (PCR), a novel Δ9- fatty acid desaturase gene...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science
2016
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| Online Access: | http://psasir.upm.edu.my/id/eprint/53322/1/Molecular%20Cloning%20and%20Functional%20Expression.pdf |
| _version_ | 1825930831222276096 |
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| author | Garba, Lawal Mohamad Ali, Mohd Shukuri Oslan, Siti Nurbaya Raja Abdul Rahman, Raja Noor Zaliha |
| author_facet | Garba, Lawal Mohamad Ali, Mohd Shukuri Oslan, Siti Nurbaya Raja Abdul Rahman, Raja Noor Zaliha |
| author_sort | Garba, Lawal |
| collection | UPM |
| description | Fatty acid desaturase enzymes play an essential role in the synthesis of unsaturated fatty acids. Pseudomonas sp. A3 was found to produce a large amount of palmitoleic and oleic acids after incubation at low temperatures. Using polymerase Chain Reaction (PCR), a novel Δ9- fatty acid desaturase gene was isolated, cloned, and successfully expressed in Escherichia coli. The gene was designated as PA3FAD9 and has an open reading frame of 1,185 bp which codes for 394 amino acids with a predicted molecular weight of 45 kDa. The activity of the gene product was confirmed via GCMS, which showed a functional putative Δ9-fatty acid desaturase capable of increasing the total amount of cellular unsaturated fatty acids of the E. coli cells expressing the gene. The results demonstrate that the cellular palmitoleic acids have increased two-fold upon expression at 15°C using only 0.1 mM IPTG. Therefore, PA3FAD9 from Pseudomonas sp.A3 codes for a Δ9-fatty acid desaturase-like protein which was actively expressed in E. coli. |
| first_indexed | 2024-03-06T09:17:32Z |
| format | Article |
| id | upm.eprints-53322 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:17:32Z |
| publishDate | 2016 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | upm.eprints-533222019-10-01T00:57:33Z http://psasir.upm.edu.my/id/eprint/53322/ Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3 Garba, Lawal Mohamad Ali, Mohd Shukuri Oslan, Siti Nurbaya Raja Abdul Rahman, Raja Noor Zaliha Fatty acid desaturase enzymes play an essential role in the synthesis of unsaturated fatty acids. Pseudomonas sp. A3 was found to produce a large amount of palmitoleic and oleic acids after incubation at low temperatures. Using polymerase Chain Reaction (PCR), a novel Δ9- fatty acid desaturase gene was isolated, cloned, and successfully expressed in Escherichia coli. The gene was designated as PA3FAD9 and has an open reading frame of 1,185 bp which codes for 394 amino acids with a predicted molecular weight of 45 kDa. The activity of the gene product was confirmed via GCMS, which showed a functional putative Δ9-fatty acid desaturase capable of increasing the total amount of cellular unsaturated fatty acids of the E. coli cells expressing the gene. The results demonstrate that the cellular palmitoleic acids have increased two-fold upon expression at 15°C using only 0.1 mM IPTG. Therefore, PA3FAD9 from Pseudomonas sp.A3 codes for a Δ9-fatty acid desaturase-like protein which was actively expressed in E. coli. Public Library of Science 2016-08 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/53322/1/Molecular%20Cloning%20and%20Functional%20Expression.pdf Garba, Lawal and Mohamad Ali, Mohd Shukuri and Oslan, Siti Nurbaya and Raja Abdul Rahman, Raja Noor Zaliha (2016) Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3. PLOS ONE, 11 (8). art. no. e0160681. pp. 1-17. ISSN 1932-6203 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0160681 10.1371/journal.pone.0160681 |
| spellingShingle | Garba, Lawal Mohamad Ali, Mohd Shukuri Oslan, Siti Nurbaya Raja Abdul Rahman, Raja Noor Zaliha Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3 |
| title | Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3 |
| title_full | Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3 |
| title_fullStr | Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3 |
| title_full_unstemmed | Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3 |
| title_short | Molecular cloning and functional expression of a Δ9-fatty acid desaturase from an antarctic Pseudomonas sp. A3 |
| title_sort | molecular cloning and functional expression of a δ9 fatty acid desaturase from an antarctic pseudomonas sp a3 |
| url | http://psasir.upm.edu.my/id/eprint/53322/1/Molecular%20Cloning%20and%20Functional%20Expression.pdf |
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