title: Engineered high content of ricinoleic acid in fission yeast Schizosaccharomyces pombe.
abstract: 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.
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology, AIST Tsukuba Central 6, Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Roman
authors.initials: R
authors.lastname: Holic
authors.forename: Hisashi
authors.initials: H
authors.lastname: Yazawa
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Kumagai
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Uemura
mesh_terms: D001709:Biotechnology / Q000379:methods; D002967:Claviceps / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D003470:Culture Media; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering / Q000379:methods; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D019301:Oleic Acid / Q000378:metabolism; D010940:Plant Proteins; D010957:Plasmids / Q000235:genetics; D012277:Ricinoleic Acids / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D013696:Temperature
chemical_list: D003470:Culture Media; D010940:Plant Proteins; D012277:Ricinoleic Acids; D019301:Oleic Acid; D006899:Mixed Function Oxygenases; C094812:phosphatidylcholine 12-monooxygenase; C030521:ricinoleic acid