Abstract
BackgroundThe ∆6-desaturase gene, encoding a key enzyme in the biosynthesis of polyunsaturated fatty acids, has potential in pharmaceutical and nutraceutical applications.ResultsThe ∆6-desaturase gene has been isolated from a selected strain of Oomycetes, Pythium sp. BCC53698. The cloned gene (PyDes6) contained an open reading frame (ORF) of 1401 bp encoding 466 amino acid residues. The deduced amino acid sequence shared a high similarity to those of other ∆6-desaturases that contained the signature features of a membrane-bound ∆6-desaturase, including a cytochrome b5 and three histidine-rich motifs and membrane-spanning regions. Heterologous expression in Saccharomyces cerevisiae showed that monoene, diene and triene fatty acids having ∆9-double bond were substrates for PyDes6. No distinct preference between the n-3 and n-6 polyunsaturated fatty acyl substrates was found. The ∆6-desaturated products were markedly increased by codon optimization of PyDes6.ConclusionThe codon-optimized ∆6-desaturase gene generated in this study is a promising tool for further reconstitution of the fatty acid profile, in a host system of choice, for the production of economically important fatty acids, particularly the n-3 and n-6 polyunsaturated fatty acids.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-015-0200-6) contains supplementary material, which is available to authorized users.
Highlights
The Δ6-desaturase gene, encoding a key enzyme in the biosynthesis of polyunsaturated fatty acids, has potential in pharmaceutical and nutraceutical applications
Polyunsaturated fatty acids (PUFAs) biosynthesis is generally associated with a set of membrane-bound enzymes, namely fatty acid desaturase and elongase, which catalyze the introduction of a double bond and the 2-carbon chain extension of fatty acids, respectively
Δ6-Desaturase is a key enzyme involved in the biosynthesis of n-3 and n-6 PUFAs, which is responsible for the conversion of essential fatty acids, linoleic acid (LA, C18:2Δ9,12) and α-linolenic acid (ALA, C18:3 Δ9,12,15) to γ-linolenic acid (GLA, C18:3Δ6,9,12) and stearidonic acid (STA, C18:4Δ6,9,12,15), respectively
Summary
The Δ6-desaturase gene, encoding a key enzyme in the biosynthesis of polyunsaturated fatty acids, has potential in pharmaceutical and nutraceutical applications. PUFA biosynthesis is generally associated with a set of membrane-bound enzymes, namely fatty acid desaturase and elongase, which catalyze the introduction of a double bond and the 2-carbon chain extension of fatty acids, respectively. These enzymes have different specificities on fatty acid substrates, relating to acyl chain length and double bond position [7]. To manipulate the oil composition of organisms of choice, by a metabolic engineering approach, a high expression level of the Δ6-desaturase enzyme in a heterologous host is required. Genes encoding for Δ6-desaturase have been cloned and characterized from various organisms [14,15,16,17,18,19,20], research on codon optimization has been limited [13]
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