Numerous Δ12-, Δ15- and multifunctional membrane fatty acid desaturases (FADs) have been identified in fungi, revealing great variability in the enzymatic specificities of FADs involved in biosynthesis of polyunsaturated fatty acids (PUFAs). Here, we report gene isolation and characterization of novel Δ12/Δ15- and Δ15-FADs named CpFad2 and CpFad3, respectively, from the opportunistic pathogenic yeast Candida parapsilosis. Overexpression of CpFad3 in Saccharomyces cerevisiae strains supplemented with linoleic acid (Δ9,Δ12-18:2) and hexadecadienoic acid (Δ9,Δ12-16:2) leads to accumulation of Δ15-PUFAs, i.e., α-linolenic acid (Δ9,Δ12,Δ15-18:3) and hexadecatrienoic acid with an unusual terminal double bond (Δ9,Δ12,Δ15-16:3). CpFad2 produces a range of Δ12- and Δ15-PUFAs. The major products of CpFad2 are linoleic and hexadecadienoic acid (Δ9,Δ12-16:2), accompanied by α-linolenic acid and hexadecatrienoic acid (Δ9,Δ12,Δ15-16:3). Using GC/MS analysis of trimethylsilyl derivatives, we identified ricinoleic acid (12-hydroxy-9-octadecenoic acid) as an additional product of CpFad2. These results demonstrate that CpFAD2 is a multifunctional FAD and indicate that detailed analysis of fatty acid derivatives might uncover a range of enzymatic selectivities in other Δ12-FADs from budding yeasts (Ascomycota: Saccharomycotina).

• MeSH
• Candida metabolismus   MeSH
• desaturasy mastných kyselin metabolismus   MeSH
• fylogeneze MeSH
• kyselina linolová metabolismus   MeSH
• kyseliny ricinolejové metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• nenasycené mastné kyseliny metabolismus   MeSH
• Saccharomyces cerevisiae metabolismus   MeSH
• sekvence aminokyselin MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• desaturasy mastných kyselin MeSH
• kyselina linolová MeSH
• kyseliny ricinolejové MeSH
• nenasycené mastné kyseliny MeSH
• ricinoleic acid MeSH Prohlížeč

The submerged culture of the Claviceps purpurea strain studied was polymorphous. In the process of alkaloid synthesis, cytodifferentiation preceded biochemical differentiation. The onset of the alkaloid phase was characterized by: predominance of chlamydospores in the culture, the presence of vegetative cells with reduced or arrested proliferation, maximum acetylCoA carboxylase activity, the maximum amount of total fatty acids, an over-average cell pool tryptophan level and minimum tryptophan synthetase activity. Intracellular ricinoleic acid was an indicator of differentiation of the culture towards alkaloid formation and also of alkaloid synthesis. The cytodifferentiation period in the initial phases of fermentation, when the cell has several alternative possibilities of development, is regarded as the most sensitive sector of the regulatory mechanisms of alkaloid formation.

• MeSH
• acetyl-CoA-karboxylasa metabolismus   MeSH
• Claviceps enzymologie   růst a vývoj   metabolismus   MeSH
• fermentace MeSH
• kyseliny ricinolejové biosyntéza   MeSH
• mastné kyseliny biosyntéza   MeSH
• námelové alkaloidy biosyntéza   MeSH
• spory hub enzymologie   růst a vývoj   metabolismus   MeSH
• tryptofan biosyntéza   MeSH
• tryptofansynthasa metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetyl-CoA-karboxylasa MeSH
• kyseliny ricinolejové MeSH
• mastné kyseliny MeSH
• námelové alkaloidy MeSH
• tryptofan MeSH
• tryptofansynthasa MeSH