Desaturation Of Palmitate Research Articles

CDNA cloning of acyl-CoA desaturase homologs in the silkworm, Bombyx mori.

We have isolated two acyl-CoA desaturase clones from a pheromone gland cDNA library by using the EST (expressed sequence tag) database of Bombyx mori. The putative acyl-CoA desaturases encoded by the clones desat 1 (2029bp) and desat 2 (2341bp) have 98% identity, and both proteins show 61% identities to Trichoplusia ni acyl-CoA Delta(11) desaturase. The deduced amino acid sequences conserve well the histidine clusters that are catalytically essential for acyl-CoA desaturase activity. Northern blot and RT-PCR analyses revealed that both transcripts of desat 1 and desat 2 were expressed predominantly in the pheromone gland. Both transcripts detected 3days before adult eclosion dramatically increased a day before adult eclosion, keeping the mRNA levels high even after eclosion. These results, combined with the fact that Delta(11) and Delta(10, 12) desaturation of palmitate is a key step to synthesize pheromone in B. mori, suggest that the desaturases encoded by desat 1 and desat 2 are involved in either or both of the desaturation steps in the pheromone biosynthetic pathway of B. mori. The mRNA levels of desat 1 and desat 2 were not affected by decapitation or injection of the pheromone biosynthesis activating neuropeptide (PBAN) into the adult female moth, suggesting that the transcription of desat 1 and desat 2 is not regulated by PBAN. In addition to the clones in the pheromone gland, eight other clones encoding the same Delta(9) desaturase homolog were found in an embryonic cDNA library by searching from the EST database of B. mori. The deduced amino acid sequence from one of the clones (desat 3) shows 79% identity to T. ni Delta(9) desaturase but only 52% identity to the desaturases in the pheromone gland of B. mori. Northern blot analysis showed that the mRNA corresponding to the desat 3 was detected in the ovary and fat body, but not in the pheromone gland. Abundance of the Delta(9) desaturase clones (eight out of the 762 randomly sequenced clones) in the library prepared from diapause-destined embryos (40h after oviposition) suggests that the Delta(9) desaturase encoded by desat 3 plays an important role in embryonic development in B. mori.

Linoleate and alpha-linolenate synthesis by isolated spinach (Spinacia oleracea) chloroplasts.

Diacylgalactosylglycerol synthesis was a prerequisite for the incorporation of [1-14C]-acetate into linoleate and alpha-linolenate of isolated spinach (Spinacia oleracea) chloroplasts. Oleate at position 1 of diacylgalactosylglycerol was desaturated to linoleate and alpha-linolenate both in the light and in the dark. Some desaturation of palmitate was also observed after prolonged incubations.

Desaturation of palmitate and stearate by cell-free fractions from soybean cotyledons.

AbstractHomogenates of cotyledons from immature soybean seeds were fractionated by centrifugation. Cell‐free preparations actively desaturated 1‐14C‐palmitate and 1‐14C‐stearate to produce 9,10‐unsaturated acids. The 9‐desaturase activity was present mainly in supernatant fractions (22,000 and 105,000 xg). These fractions also desaturated oleate to linoleate and elongated C14, C16 and C18 acids. In view of this versatility of desaturase systems in the soybean, including the 9‐desaturase(s) for C14, C16 and C18 saturated acids, there would not seem to be further need to consider a separate plant pathway for biosynthesis of unsaturated fatty acids.