Naslov
Plinska kromatografija masnih kiselina iz kvasca transformiranog genom za elongazu masnih kiselina (kcs) iz biljaka
(Gas chromatography of fatty acids in transformed yeast upon induced expression of a plant fatty acid elongase (KCS))

Autori
Vrdoljak, Marijana ; Posavec, Rudolf ; Mihaljević, Snježana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, ostalo

Izvornik
Book of Abstracts / Martinez, Sanja - Zagreb : Hrvatsko društvo kemijskih inženjera i tehnologa (HDKI), 2012, 74-74

ISBN
987-953-6894-45-1

Skup
IX. Susret mladih kemijskih inženjera - Nove tehnologije i transfer znanja

Mjesto i datum
Zagreb, Hrvatska, 16.02.2012. - 17.02.2012

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
ekspresija; gen KCS; kvasac Saccharomyces cerevisiae; nervonska kiselina; plinska kromatografija
(expression; gas-chromatography; gene KCS; nervonic acid; yeast Saccharomyces cerevisiae)

Sažetak
The 3-ketoacyl-CoA synthase (KCS ; EC 2.3.1.86) is a fatty acid elongase, which catalyzes the synthesis of several very long chain monounsaturated fatty acids (VLCMFA) in plants. The aim of this study was to clone KCS gene (CgKCS) from plant species Cardamine graeca L. into yeast cells and to analyse expression of the recombinant protein. Coding region of the CgKCS was linked to the galactose-inducible GAL1 promoter in the expression vector pYES2 and introduced into Saccharomyces cerevisiae strain BY4741. After the induced expression, total fatty acid content and acyl composition in transformed yeast was determined by preparing fatty acid methyl esters (FAME) extracted in hexane and analyzed by gas-chromatography (GC) using the 37 Component FAME Mix (Supelco) as an external standard. The conditions used were optimized to allow the detection of VLCFAs of up to C24 in length. Separation of FAMEs was affected by a type of column (HP-5 or DB-FFAP, Agilent)) and temperature program rate. Comparative GC analyses of fatty acid profiles in transformed yeast cells upon CgKCS expression revealed the presence of saturated and monounsaturated VLCFAs that are not normally present in untransformed yeast, with a distinct peak corresponding to nervonic acid 24:1 Δ15 (cis-tetracos-15-enoic acid).

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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