title: Heterologous Expression of PA8FAD9 and Functional Characterization of a Δ9-Fatty Acid Desaturase from a Cold-Tolerant Pseudomonas sp. A8.
abstract: 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.
authors.affiliation: Enzyme and Microbial Technology Research Centre, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia.|Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia.|Department of Microbiology, Faculty of Science, Gombe State University, Tudun Wada Gombe, P.M.B 127, Gombe State, Nigeria.
authors.forename: Lawal
authors.initials: L
authors.lastname: Garba
authors.affiliation: Enzyme and Microbial Technology Research Centre, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia.|Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia.
authors.forename: Mohd Shukuri Mohamad
authors.initials: MS
authors.lastname: Ali
authors.affiliation: Enzyme and Microbial Technology Research Centre, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia.|Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia.
authors.forename: Siti Nurbaya
authors.initials: SN
authors.lastname: Oslan
authors.affiliation: Enzyme and Microbial Technology Research Centre, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia. rnzaliha@upm.edu.my.|Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia. rnzaliha@upm.edu.my.
authors.forename: Raja Noor Zaliha Raja Abdul
authors.initials: RN
authors.lastname: Rahman
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D003080:Cold Temperature; D004926:Escherichia coli / Q000235:genetics; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism*; D005229:Fatty Acids, Monounsaturated / Q000378:metabolism; D008970:Molecular Weight; D016366:Open Reading Frames; D019308:Palmitic Acid / Q000378:metabolism; D010802:Phylogeny; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D013229:Stearic Acids / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005229:Fatty Acids, Monounsaturated; D013229:Stearic Acids; C008757:palmitoleic acid; D019308:Palmitic Acid; C031183:stearic acid; D044943:Fatty Acid Desaturases
keywords: Expression; Membrane phospholipids; Pseudomonas sp. A8; Unsaturated fatty acids; Δ9-Fatty acid desaturase