Open AccessMolecular Cloning and Functional Expression of a Δ9- Fatty Acid Desaturase from an Antarctic Pseudomonas sp. A3
Author(s) -
Lawal Garba,
Mohd Shukuri Mohamad Ali,
Siti Nurbaya Oslan,
Raja Noor Zaliha Raja Abdul Rahman
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0160681
Subject(s) - biochemistry , palmitoleic acid , escherichia coli , biology , fatty acid desaturase , fatty acid , stearoyl coa desaturase , pseudomonas , oleic acid , gene , open reading frame , enzyme , molecular cloning , cloning (programming) , gene expression , bacteria , polyunsaturated fatty acid , peptide sequence , linoleic acid , genetics , computer science , programming language
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 .