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Engineered high content of ricinoleic acid in fission yeast Schizosaccharomyces pombe

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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.

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Acknowledgments

A part of this work was supported by the A-STEP (Adaptable and Seamless Technology transfer Program through target-driven R&D) research grant from Japan Science an122222d Technology Agency (JST) to HK and HU. RH was a recipient of “JSPS Postdoctoral Fellowship for Foreign Researchers” from the Japan Society for the Promotion of Science. We wish to thank Masakazu Yamaoka, Yasushi Kamisaka, and Kazuyoshi Kimura for continuous encouragement.

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  1. Roman Holic

    Present address: Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Moyzesova 61, 900 28, Ivanka pri Dunaji, Bratislava, Slovakia

Authors and Affiliations

  1. Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology, AIST Tsukuba Central 6, Higashi 1-1-1, Tsukuba, Ibaraki, 305-8566, Japan

    Roman Holic, Hisashi Yazawa & Hiroshi Uemura

  2. ASPEX Division, Asahi Glass Co. Ltd, 1-5-1 Marunouchi, Chiyoda-ku, Tokyo, 100-8405, Japan

    Hiromichi Kumagai

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  1. Roman Holic
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  2. Hisashi Yazawa
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Correspondence to Hiroshi Uemura.

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Holic, R., Yazawa, H., Kumagai, H. et al. Engineered high content of ricinoleic acid in fission yeast Schizosaccharomyces pombe . Appl Microbiol Biotechnol 95, 179–187 (2012). https://doi.org/10.1007/s00253-012-3959-6

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