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Applied and Environmental Microbiology, November 2007, p. 6965-6971, Vol. 73, No. 21
0099-2240/07/$08.00+0     doi:10.1128/AEM.01008-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Heterologous Production of Dihomo-{gamma}-Linolenic Acid in Saccharomyces cerevisiae{triangledown}

Hisashi Yazawa,1 Hitoshi Iwahashi,1 Yasushi Kamisaka,1 Kazuyoshi Kimura,1 Tsunehiro Aki,2 Kazuhisa Ono,2 and Hiroshi Uemura1*

National Institute of Advanced Industrial Science and Technology, AIST Tsukuba Central 6, Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan,1 Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8530, Japan2

Received 5 May 2007/ Accepted 3 September 2007

To make dihomo-{gamma}-linolenic acid (DGLA) (20:3n-6) in Saccharomyces cerevisiae, we introduced Kluyveromyces lactis {Delta}12 fatty acid desaturase, rat {Delta}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 µg/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.

* Corresponding author. Mailing address: Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, AIST Tsukuba Central 6, Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan. Phone: 81-298-61-6425. Fax: 81-298-61-6172. E-mail: hiroshi-uemura{at}aist.go.jp

{triangledown} Published ahead of print on 14 September 2007.

Applied and Environmental Microbiology, November 2007, p. 6965-6971, Vol. 73, No. 21
0099-2240/07/$08.00+0     doi:10.1128/AEM.01008-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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