Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8
Fatty acid desaturase enzymes are capable of inserting double bonds between carbon atoms of saturated fatty acyl-chains to produce unsaturated fatty acids. A gene coding for a putative Δ9-fatty acid desaturase-like protein was isolated from a cold-tolerant Pseudomonas sp. A8, cloned and heterologous...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Humana Press
2016
|
| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/54430/1/Heterologous%20expression%20of%20PA8FAD9%20and%20functional%20characterization%20.pdf |
| _version_ | 1825931036605808640 |
|---|---|
| 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 are capable of inserting double bonds between carbon atoms of saturated fatty acyl-chains to produce unsaturated fatty acids. A gene coding for a putative Δ9-fatty acid desaturase-like protein was isolated from a cold-tolerant Pseudomonas sp. A8, cloned and heterologously expressed in Escherichia coli. The gene named as PA8FAD9 has an open reading frame of 1185 bp and codes for 394 amino acids with a predicted molecular weight of 45 kDa. The enzyme showed high Δ9-fatty acid desaturase-like protein activity and increased overall levels of cellular unsaturated fatty acids in the recombinant E. coli cells upon expression at different temperatures. The results showed that the ratio of palmitoleic to palmitic acid in the recombinant E. coli cells increased by more than twice the amount observed in the control cells at 20 °C using 0.4 mM IPTG. GCMS analysis confirmed the ability of this enzyme to convert exogenous stearic acid to oleic acid incorporated into the recombinant E. coli membrane phospholipids. It may be concluded that the PA8FAD9 gene from Pseudomonas sp. A8 codes for a putative Δ9-fatty acid desaturase protein actively expressed in E. coli under the influence of temperature and an inducer. |
| first_indexed | 2024-03-06T09:20:39Z |
| format | Article |
| id | upm.eprints-54430 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:20:39Z |
| publishDate | 2016 |
| publisher | Humana Press |
| record_format | dspace |
| spelling | upm.eprints-544302018-03-16T02:40:56Z http://psasir.upm.edu.my/id/eprint/54430/ Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8 Garba, Lawal Mohamad Ali, Mohd Shukuri Oslan, Siti Nurbaya Raja Abdul Rahman, Raja Noor Zaliha Fatty acid desaturase enzymes are capable of inserting double bonds between carbon atoms of saturated fatty acyl-chains to produce unsaturated fatty acids. A gene coding for a putative Δ9-fatty acid desaturase-like protein was isolated from a cold-tolerant Pseudomonas sp. A8, cloned and heterologously expressed in Escherichia coli. The gene named as PA8FAD9 has an open reading frame of 1185 bp and codes for 394 amino acids with a predicted molecular weight of 45 kDa. The enzyme showed high Δ9-fatty acid desaturase-like protein activity and increased overall levels of cellular unsaturated fatty acids in the recombinant E. coli cells upon expression at different temperatures. The results showed that the ratio of palmitoleic to palmitic acid in the recombinant E. coli cells increased by more than twice the amount observed in the control cells at 20 °C using 0.4 mM IPTG. GCMS analysis confirmed the ability of this enzyme to convert exogenous stearic acid to oleic acid incorporated into the recombinant E. coli membrane phospholipids. It may be concluded that the PA8FAD9 gene from Pseudomonas sp. A8 codes for a putative Δ9-fatty acid desaturase protein actively expressed in E. coli under the influence of temperature and an inducer. Humana Press 2016-11 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/54430/1/Heterologous%20expression%20of%20PA8FAD9%20and%20functional%20characterization%20.pdf Garba, Lawal and Mohamad Ali, Mohd Shukuri and Oslan, Siti Nurbaya and Raja Abdul Rahman, Raja Noor Zaliha (2016) Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8. Molecular Biotechnology, 58 (11). pp. 718-728. ISSN 1073-6085; ESSN: 1559-0305 https://link.springer.com/article/10.1007%2Fs12033-016-9971-9 Pseudomonas sp. A8; Δ9-fatty acid desaturase; Unsaturated fatty acids; Expression; Membrane phospholipids 10.1007/s12033-016-9971-9 |
| spellingShingle | Pseudomonas sp. A8; Δ9-fatty acid desaturase; Unsaturated fatty acids; Expression; Membrane phospholipids Garba, Lawal Mohamad Ali, Mohd Shukuri Oslan, Siti Nurbaya Raja Abdul Rahman, Raja Noor Zaliha Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8 |
| title | Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8 |
| title_full | Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8 |
| title_fullStr | Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8 |
| title_full_unstemmed | Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8 |
| title_short | Heterologous expression of PA8FAD9 and functional characterization of a Δ9-fatty acid desaturase from a cold-tolerant Pseudomonas sp. A8 |
| title_sort | heterologous expression of pa8fad9 and functional characterization of a δ9 fatty acid desaturase from a cold tolerant pseudomonas sp a8 |
| topic | Pseudomonas sp. A8; Δ9-fatty acid desaturase; Unsaturated fatty acids; Expression; Membrane phospholipids |
| url | http://psasir.upm.edu.my/id/eprint/54430/1/Heterologous%20expression%20of%20PA8FAD9%20and%20functional%20characterization%20.pdf |
| work_keys_str_mv | AT garbalawal heterologousexpressionofpa8fad9andfunctionalcharacterizationofad9fattyaciddesaturasefromacoldtolerantpseudomonasspa8 AT mohamadalimohdshukuri heterologousexpressionofpa8fad9andfunctionalcharacterizationofad9fattyaciddesaturasefromacoldtolerantpseudomonasspa8 AT oslansitinurbaya heterologousexpressionofpa8fad9andfunctionalcharacterizationofad9fattyaciddesaturasefromacoldtolerantpseudomonasspa8 AT rajaabdulrahmanrajanoorzaliha heterologousexpressionofpa8fad9andfunctionalcharacterizationofad9fattyaciddesaturasefromacoldtolerantpseudomonasspa8 |