Functional characterization and overexpression of Δ12-desaturase in the oleaginous yeast Rhodotorula toruloides for production of linoleic acid-rich lipids

2021 ◽  
Author(s):  
Chih-Chan Wu ◽  
Takao Ohashi ◽  
Hiroyuki Kajiura ◽  
Yu Sato ◽  
Ryo Misaki ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Oleaginous Yeast ◽  
Functional Characterization ◽  
Δ12 Desaturase

Saccharomyces kluyveri FAD3 encodes an ω3 fatty acid desaturase

Microbiology ◽  
2004 ◽  
Vol 150 (6) ◽  
pp. 1983-1990 ◽  
Author(s):  
Takahiro Oura ◽  
Susumu Kajiwara
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Linoleic Acid ◽  
Linolenic Acid ◽  
Functional Characterization ◽  
Fatty Acid Desaturase ◽  
Fatty Acid Desaturases ◽  
Desaturase Gene ◽  
Saccharomyces Kluyveri ◽  
Fatty Acid Desaturase Gene

Fungi, like plants, are capable of producing the 18-carbon polyunsaturated fatty acids linoleic acid and α-linolenic acid. These fatty acids are synthesized by catalytic reactions of Δ12 and ω3 fatty acid desaturases. This paper describes the first cloning and functional characterization of a yeast ω3 fatty acid desaturase gene. The deduced protein encoded by the Saccharomyces kluyveri FAD3 gene (Sk-FAD3) consists of 419 amino acids, and shows 30–60 % identity with Δ12 fatty acid desaturases of several eukaryotic organisms and 29–31 % identity with ω3 fatty acid desaturases of animals and plants. During Sk-FAD3 expression in Saccharomyces cerevisiae, α-linolenic acid accumulated only when linoleic acid was added to the culture medium. The disruption of Sk-FAD3 led to the disappearance of α-linolenic acid in S. kluyveri. These findings suggest that Sk-FAD3 is the only ω3 fatty acid desaturase gene in this yeast. Furthermore, transcriptional expression of Sk-FAD3 appears to be regulated by low-temperature stress in a manner different from the other fatty acid desaturase genes in S. kluyveri.

Combined transgenic expression of Δ12-desaturase and Δ12-epoxygenase in high linoleic acid seeds leads to increased accumulation of vernolic acid

2006 ◽  
Vol 33 (6) ◽  
pp. 585 ◽  
Author(s):  
Xue-Rong Zhou ◽  
Surinder Singh ◽  
Qing Liu ◽  
Allan Green
Keyword(s):  
Fatty Acids ◽  
Linoleic Acid ◽  
Vernolic Acid ◽  
Renewable Resource ◽  
High Linoleic Acid ◽  
Transgenic Expression ◽  
Unusual Fatty Acids ◽  
Δ12 Desaturase ◽  
Seed Crops ◽  
Acid Substrate

The transgenic production of unusual fatty acids in oil seed crops offers an alternative, renewable resource for industry. However, transgenic expression of genes catalysing the synthesis of unusual fatty acids has generally resulted in these fatty acids accumulating at levels significantly below the levels in the wild species from which the genes were sourced. This study reports expression of additional copies of any of three Δ12-desaturase genes (FAD2) from Crepis palaestina Bornm., cotton (Gossypium hirsutum L.) or Arabidopsis thaliana (L.) Heynh. with C. palaestina Δ12-epoxygenase gene (Cpal2), in an Arabidopsis mutant having a significantly higher level of linoleic acid substrate. This resulted in the highest levels of vernolic acid accumulation, 21% of total fatty acids, reported so far in any transgenic plant expressing the Δ12-epoxygenase. Similarly, the co-expression of C. palaestina Cpal2 and a transgenic copy of FAD2 in cotton seed that contains large amounts of linoleic acid substrate also resulted in greater accumulation of vernolic acid in seed than did expression of C. palaestina Cpal2 alone.

The molecular basis of the high linoleic acid content in Petunia seed oil: analysis of a seed-specific linoleic acid mutant

2000 ◽  
Vol 28 (6) ◽  
pp. 631-632 ◽  
Author(s):  
I. Verwoert ◽  
Y. Meller-Harel ◽  
K. van der Linden ◽  
B. Verbree ◽  
R. Koes ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Acid Content ◽  
Seed Oil ◽  
Lipid Fraction ◽  
Lipid Synthesis ◽  
Linoleic Acid Content ◽  
Specific Expression ◽  
High Linoleic Acid ◽  
High Linoleic Acid Content ◽  
Δ12 Desaturase

From a random transposon mutagenesis experiment, using Petunia line W138, a seed-specific linoleic acid mutant was isolated. The tagged gene was cloned and identified as a microsomal Δ12 desaturase. Expression of the gene, however, was constitutive and not, as might have been expected, seed-specific. Moreover, self-fertilized homozygous mutants still contain 40% 18:2 in the seed lipid fraction. This suggests that at least two (seedspecific) Δ12 desaturase genes are responsible for the high linoleic acid content in Petunia seed oil. Five members of the microsomal Δ12 desaturase gene family have been identified and isolated. Data are presented on the molecular characterization and tissue-specific expression of these genes, which suggest that, in Petunia the flux through the prokaryotic and eukaryotic pathways of lipid synthesis might be different from the situation found in Arabidopsis.

Isolation and functional characterization of two independently-evolved fatty acid Δ12-desaturase genes from insects

2008 ◽  
Vol 17 (6) ◽  
pp. 667-676 ◽  
Author(s):  
X.-R. Zhou ◽  
I. Horne ◽  
K. Damcevski ◽  
V. Haritos ◽  
A. Green ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Functional Characterization ◽  
Δ12 Desaturase

scholarly journals De novo synthesis of trans-10, cis-12 conjugated linoleic acid in oleaginous yeast Yarrowia Lipolytica

2012 ◽  
Vol 11 (1) ◽  
pp. 51 ◽  
Author(s):  
Baixi Zhang ◽  
Chunchi Rong ◽  
Haiqin Chen ◽  
Yuanda Song ◽  
Hao Zhang ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Yarrowia Lipolytica ◽  
Oleaginous Yeast ◽  
De Novo ◽  
De Novo Synthesis ◽  
Yeast Yarrowia Lipolytica

A Novel Reconstitution Method for Inducing the Formation of Regular 2-Dimensional Arrays of Membrane Proteins and Lipids

1988 ◽  
Vol 46 ◽  
pp. 152-153 ◽  
Author(s):  
A. Engel ◽  
A. Holzenburg ◽  
K. Stauffer ◽  
J. Rosenbusch ◽  
U. Aebi
Keyword(s):  
Membrane Proteins ◽  
Flow Rate ◽  
Lipid Membranes ◽  
Functional Characterization ◽  
Dimensional Structure ◽  
Electron Crystallography ◽  
Peltier Element ◽  
Protein Ratio ◽  
Precise Control ◽  
3 Dimensional

Reconstitution of solubilized and purified membrane proteins in the presence of phospholipids into vesicles allows their functions to be studied by simple bulk measurements (e.g. diffusion of differently sized solutes) or by conductance measurements after transformation into planar membranes. On the other hand, reconstitution into regular protein-lipid arrays, usually forming at a specific lipid-to-protein ratio, provides the basis for determining the 3-dimensional structure of membrane proteins employing the tools of electron crystallography.To refine reconstitution conditions for reproducibly inducing formation of large and highly ordered protein-lipid membranes that are suitable for both electron crystallography and patch clamping experiments aimed at their functional characterization, we built a flow-dialysis device that allows precise control of temperature and flow-rate (Fig. 1). The flow rate is generated by a peristaltic pump and can be adjusted from 1 to 500 ml/h. The dialysis buffer is brought to a preselected temperature during its travel through a meandering path before it enters the dialysis reservoir. A Z-80 based computer controls a Peltier element allowing the temperature profile to be programmed as function of time.

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