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Volume 145, Issue 10

Two fatty acid Δ9-desaturase genes, and , from complement the yeast mutation Free

Abstract

Genes encoding two distinct fatty acid Δ9-desaturases were isolated from strains of the oleaginous fungus . Two genomic sequences, Δ9-1 and Δ9-2, each containing a single intron, were cloned from strain CBS 528.72 while one cDNA clone, LM9, was isolated from strain CBS 210.32. The Δ9-1 gene encoded a protein of 445 aa which shared 99% identity with the LM9 gene product. These proteins also showed 40–60% identity to the Δ9-desaturases (Ole1p) of other fungi and contained the three conserved histidine boxes, C-terminal cytochrome fusion and transmembrane domains characteristic of endoplasmic reticulum membrane-bound Δ9-desaturases. LM9 and Δ9-1 are therefore considered to represent the same gene (). The gene was transcriptionally active in all strains tested and its function was confirmed by complementation of the mutation. Fatty acid analysis of yeast transformants expressing the CBS 210.32 gene showed an elevated level of oleic acid (18:1) compared to palmitoleic acid (16:1), the major fatty acid component of wild-type . This indicated that the Δ9-desaturase had a substrate preference for stearic acid (18:0) rather than palmitic acid (16:0). Genomic clone Δ9-2 () also encoded a protein of 445 aa which had 86% identity to the Δ9-1 and LM9 proteins and whose ORF also complemented the yeast mutation. The transcript from this gene could only be detected in one of the six strains tested, suggesting that its expression may be strain-specific or induced under certain physiological conditions.

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Keyword(s): 16:0, palmitic acid , 16:1, palmitoleic acid , 18:0, stearic acid , 18:1, oleic acid , 18:2, linoleic acid , 18:3, α-linolenic acid , 20:3, dihomo-γ-linolenic acid , 20:4, arachidonic acid (ARA) , ER, endoplasmic reticulum , fatty acid desaturation , LCPUFA, long-chain polyunsaturated fatty acid , Mortierella alpina , oleaginous fungus , RACE, rapid amplification of cDNA ends , UTR, untranslated region , yeast complementation , γ-18:3, γ-linolenic acid and Δ9-desaturase genes
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1999-10-01
2024-05-03
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Two fatty acid Δ9-desaturase genes, ole1 and ole2, from Mortierella alpina complement the yeast ole1 mutation
Microbiology 145, 2939 (1999); https://doi.org/10.1099/00221287-145-10-2939
/content/journal/micro/10.1099/00221287-145-10-2939
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