Identification of Genes Affecting Lipid Content Using Transposon Mutagenesis in<i>Saccharomyces cerevisiae</i>

Kamisaka, Yasushi; Noda, Naomi; Tomita, Nao; Kimura, Kouji; Kodaki, Tsutomu; Hosaka, K.

doi:10.1271/bbb.70.646

Cited by 42 publications

(33 citation statements)

References 38 publications

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“…However, total lipid content in S. pombe cell is not high and around 5% to 10% of dry cell weight (DCW). This value is similar to the level in S. cerevisiae (Kamisaka et al 2006). For instance, the total FA content/DCW of sample C in Fig.…”

Section: Discussionsupporting

confidence: 84%

Engineered high content of ricinoleic acid in fission yeast Schizosaccharomyces pombe

Holic̆

Yazawa

Kumagai

et al. 2012

Appl Microbiol Biotechnol

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In an effort to produce ricinoleic acid (12-hydroxy-octadeca-cis-9-enoic acid: C18:1-OH) as a petrochemical replacement in a variety of industrial processes, we introduced Claviceps purpurea oleate ∆12-hydroxylase gene (CpFAH12) to Schizosaccharomyces pombe, putting it under the control of inducible nmt1 promoter. Since Fah12p is able to convert oleic acid to ricinoleic acid, we thought that S. pombe, in which around 75% of total fatty acid (FA) is oleic acid, would accordingly be an ideal microorganism for high production of ricinoleic acid. Unfortunately, at the normal growth temperature of 30 °C, S. pombe cells harboring CpFAH12 grew poorly when the CpFAH12 gene expression was induced, perhaps implicating ricinoleic acid as toxic in S. pombe. However, in line with a likely thermoinstability of Fah12p, there was almost no growth inhibition at 37 °C or, by contrast with 30 °C and lower temperatures, ricinoleic acid accumulation. Accordingly, various optimization steps led to a regime with preliminary growth at 37 °C followed by a 5-day incubation at 20 °C, and the level of ricinoleic acid reached 137.4 μg/ml of culture that corresponded to 52.6% of total FA.

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Section: Discussionsupporting

confidence: 84%

Engineered high content of ricinoleic acid in fission yeast Schizosaccharomyces pombe

Holic̆

Yazawa

Kumagai

et al. 2012

Appl Microbiol Biotechnol

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“…3% of fatty acid content, comparable to the values obtained in the systems of Beaudoin et al (2) and Domergue et al (8). The total lipid content of S. cerevisiae is not high (22) and may have to be increased for our long-term goal of producing PUFAs by use of transgenic S. cerevisiae in sustainable quantities.…”

Section: (0)supporting

confidence: 78%

“…NSD was examined because the carbon/nitrogen ratio affects the amount of lipids in the cell (22,32). Prior to this experiment, we preliminarily examined the effect of carbon sources that result in oxidative metabolism, as PUFA production requires an active cytochrome system.…”

Section: Resultsmentioning

confidence: 99%

Heterologous Production of Dihomo-γ-Linolenic Acid in Saccharomyces cerevisiae

Yazawa

Iwahashi

Kamisaka

et al. 2007

Appl Environ Microbiol

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To make dihomo-␥-linolenic acid (DGLA) (20:3n-6) in Saccharomyces cerevisiae, we introduced Kluyveromyces lactis ⌬12 fatty acid desaturase, rat ⌬6 fatty acid desaturase, and rat elongase genes. Because Fad2p is able to convert the endogenous oleic acid to linoleic acid, this allowed DGLA biosynthesis without the need to supply exogenous fatty acids on the media. Medium composition, cultivation temperature, and incubation time were examined to improve the yield of DGLA. Fatty acid content was increased by changing the medium from a standard synthetic dropout medium to a nitrogen-limited minimal medium (NSD). Production of DGLA was higher in the cells grown at 15°C than in those grown at 20°C, and no DGLA production was observed in the cells grown at 30°C. In NSD at 15°C, fatty acid content increased up until day 7 and decreased after day 10. When the cells were grown in NSD for 7 days at 15°C, the yield of DGLA reached 2.19 g/mg of cells (dry weight) and the composition of DGLA to total fatty acids was 2.74%. To our knowledge, this is the first report describing the production of polyunsaturated fatty acids in S. cerevisiae without supplying the exogenous fatty acids.

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“…TAG metabolism is well characterized in S. cerevisiae (Sorger and Daum 2003;Czabany et al 2007), and overexpression of DGAT genes has been reported to increase the lipid content in plants and yeasts (Hobbs et al 1999;Bouvier-Nave et al 2000;Jako et al 2001). Kamisaka et al (2006) found that the disruption of SNF2, a gene encoding a transcription factor forming part of the SWI/SNF chromatin-remodeling complex, increased the lipid content in S. cerevisiae. Overexpression of DGA1 (diacylglycerol acyltransferase) and LEU2 genes in the snf2 disruptant has achieved 29 % of total lipid content to DCW, high enough to use this strain as a model of oleaginous yeast (Kamisaka et al 2007).…”

Section: Future Perspectivesmentioning

confidence: 99%

Synthesis and production of unsaturated and polyunsaturated fatty acids in yeast: current state and perspectives

Uemura

2012

Appl Microbiol Biotechnol

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Recently, many genes involved in the formation of unsaturated and polyunsaturated fatty acids (PUFAs) were isolated. In most cases, their activities were confirmed by expressing them in the well-studied model organism Saccharomyces cerevisiae because its fatty acid compositions are very simple and it does not contain PUFAs. Taking advantage of its genetic tractability and increasing wealth of accessible data, many groups are attempting to produce various useful fatty acids in the model yeasts, mainly in S. cerevisiae. This review describes typical such examples including a very recent study on the expression of a fatty acid hydroxylase gene in fission yeast Schizosaccharomyces pombe. Furthermore, the impact of the genetically engineered alteration of fatty acid composition on the stress tolerance is presented because unsaturated fatty acids have crucial roles in membrane fluidity and signaling processes. Lastly, recent attempts at increasing lipid content in S. cerevisiae are discussed.

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Identification of Genes Affecting Lipid Content Using Transposon Mutagenesis inSaccharomyces cerevisiae

Cited by 42 publications

References 38 publications

Engineered high content of ricinoleic acid in fission yeast Schizosaccharomyces pombe

Engineered high content of ricinoleic acid in fission yeast Schizosaccharomyces pombe

Heterologous Production of Dihomo-γ-Linolenic Acid in Saccharomyces cerevisiae

Synthesis and production of unsaturated and polyunsaturated fatty acids in yeast: current state and perspectives

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