Identification of Δ5-Desaturase from Mortierella alpina by Heterologous Expression in Bakers' Yeast and Canola

Knutzon, Deborah S.; Thurmond, Jennifer M.; Huang, Y. S.; Chaudhary, Sunita; Bobik, E.; Chan, George M.; Kirchner, Stephen J.; Mukerji, Pradip

doi:10.1074/jbc.273.45.29360

Cited by 139 publications

(107 citation statements)

References 33 publications

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“…In clade C, a functionally characterized Δ5 Fad from the oleaginous fungus, Mortierella alpina , was found (Michaelson, Lazarus, Griffiths, Napier, & Stobart, 1998). The fatty acid profile of M. alpina has been found to have high levels of ARA and also EPA when conditions are optimal, but lack > C 20 PUFAs, such as DHA (Knutzon et al., 1998; Michaelson et al., 1998). Furthermore, M. alpina has been shown to encode an additional Δ6 desaturase enzyme; however, this sequence is not present in the Swiss‐prot database and therefore was not included in this analysis (Knutzon et al., 1998; Michaelson et al., 1998).…”

Section: Discussionmentioning

confidence: 99%

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Surm

Toledo

Prentis

et al. 2018

Ecology and Evolution

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The biosynthesis of long‐chain polyunsaturated fatty acids (LC‐PUFAs, ≥ C20) is reliant on the action of desaturase and elongase enzymes, which are encoded by the fatty acyl desaturase (Fad) and elongation of very long‐chain fatty acid (Elovl) gene families, respectively. In Metazoa, research investigating the distribution and evolution of these gene families has been restricted largely to Bilateria. Here, we provide insights into the phylogenetic and molecular evolutionary histories of the Fad and Elovl gene families in Cnidaria, the sister phylum to Bilateria. Four model cnidarian genomes and six actiniarian transcriptomes were interrogated. Analysis of the fatty acid composition of a candidate cnidarian species, Actinia tenebrosa, was performed to determine the baseline profile of this species. Phylogenetic analysis revealed lineage‐specific gene duplication in actiniarians for both the Fad and Elovl gene families. Two distinct cnidarian Fad clades clustered with functionally characterized Δ5 and Δ6 proteins from fungal and plant species, respectively. Alternatively, only a single cnidarian Elovl clade clustered with functionally characterized Elovl proteins (Elovl4), while two additional clades were identified, one actiniarian‐specific (Novel ElovlA) and the another cnidarian‐specific (Novel ElovlB). In actiniarians, selection analyses revealed pervasive purifying selection acting on both gene families. However, codons in the Elovl gene family show patterns of nucleotide variation consistent with the action of episodic diversifying selection following gene duplication events. Significantly, these codons may encode amino acid residues that are functionally important for Elovl proteins to target and elongate different precursor fatty acids. In A. tenebrosa, the fatty acid analysis revealed an absence of LC‐PUFAs > C20 molecules and implies that the Elovl enzymes are not actively contributing to the elongation of these LC‐PUFAs. Overall, this study has revealed that actiniarians possess Fad and Elovl genes required for the biosynthesis of some LC‐PUFAs, and that these genes appear to be distinct from bilaterians.

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Section: Discussionmentioning

confidence: 99%

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Surm

Toledo

Prentis

et al. 2018

Ecology and Evolution

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“…Related Δ 5 -desaturases are known from plants, fungi, and humans, however the primary structure of JamB is distinct from each of these. The third His box domain in JamB aligns with the Δ 5 -/Δ 6 -desaturases, where the consensus sequence is QXXHH [224]. The functional significance of variations in this signature domain is not known.…”

Section: Jamaicamides-jamaicamidesmentioning

confidence: 99%

Biosynthesis and function of polyacetylenes and allied natural products

Minto

Blacklock

2008

Progress in Lipid Research

288

227

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Polyacetylenic natural products are a substantial class of often unstable compounds containing a unique carbon-carbon triple bond functionality, that are intriguing for their wide variety of biochemical and ecological functions, economic potential, and surprising mode of biosynthesis. Isotopic tracer experiments between 1960 and 1990 demonstrated that the majority of these compounds are derived from fatty acid and polyketide precursors. During the past decade, research into the metabolism of polyacetylenes has swiftly advanced, driven by the cloning of the first genes responsible for polyacetylene biosynthesis in plants, moss, fungi, and actinomycetes, and the initial characterization of the gene products.The current state of knowledge of the biochemistry and molecular genetics of polyacetylenic secondary metabolic pathways will be presented together with an up-to-date survey of new terrestrial and marine natural products, their known biological activities, and a discussion of their likely metabolic origins.

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“…1. Although the two desaturases (⌬ 6 -and ⌬ 5 -fatty acid desaturases) required for this pathway have been cloned recently from a number of different sources (7)(8)(9)(10)(11)(12)(13)(14), no genes have been characterized for the C2 elongation of the C18 PUFA. Biochemical characterization of mammalian elongation systems (most notably from liver microsomes) has indicated that the ''elongase'' actually consists of four enzyme activities, being made up of a condensing enzyme, a ␤-ketoreductase, a dehydrase, and an enoyl reductase (reviewed in ref.…”

mentioning

confidence: 99%

Heterologous reconstitution in yeast of the polyunsaturated fatty acid biosynthetic pathway

Beaudoin

Michaelson

Hey

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

144

114

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A Caenorhabditis elegans ORF encoding the presumptive condensing enzyme activity of a fatty acid elongase has been characterized functionally by heterologous expression in yeast. This ORF (F56H11.4) shows low similarity to Saccharomyces cerevisiae genes involved in fatty acid elongation. The substrate specificity of the C. elegans enzyme indicated a preference for ⌬ 6 -desaturated C18 polyunsaturated fatty acids. Coexpression of this activity with fatty acid desaturases required for the synthesis of C20 polyunsaturated fatty acids resulted in the accumulation of arachidonic acid from linoleic acid and eicosapentaenoic acid from ␣-linolenic acid. These results demonstrate the reconstitution of the n-3 and n-6 polyunsaturated fatty acid biosynthetic pathways. The C. elegans ORF is likely to interact with endogenous components of a yeast elongation system, with the heterologous nematode condensing enzyme F56H11.4 causing a redirection of enzymatic activity toward polyunsaturated C18 fatty acid substrates.

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Identification of Δ5-Desaturase from Mortierella alpina by Heterologous Expression in Bakers' Yeast and Canola

Cited by 139 publications

References 33 publications

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Insights into the phylogenetic and molecular evolutionary histories of Fad and Elovl gene families in Actiniaria

Biosynthesis and function of polyacetylenes and allied natural products

Heterologous reconstitution in yeast of the polyunsaturated fatty acid biosynthetic pathway

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