Suppression of ricinoleic acid toxicity by ptl2 overexpression in fission yeast Schizosaccharomyces pombe
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- 作者：Hisashi Yazawa (1)
Masayo Ogiso (1)
Hiromichi Kumagai (2)
Hiroshi Uemura (1) - 关键词：Schizosaccharomyces pombe ; Yeast ; Ricinoleic acid ; FAH12 ; Fatty acid hydroxylase ; Lipase
- 刊名：Applied Microbiology and Biotechnology
- 出版年：2014
- 出版时间：November 2014
- 年：2014
- 卷：98
- 期：22
- 页码：9325-9337
- 全文大小：811 KB
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Masayo Ogiso (1)
Hiromichi Kumagai (2)
Hiroshi Uemura (1)
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
2. Asahi Glass Co. Ltd., Yokohama, Kanagawa, 221-8755, Japan - ISSN：1432-0614
文摘
We previously succeeded to obtain a high content of ricinoleic acid (RA), a hydroxylated fatty acid with great values as a petrochemical replacement, in fission yeast Schizosaccharomyces pombe by introducing Claviceps purpurea oleate Δ12-hydroxylase gene (CpFAH12). Although the production was toxic to S. pombe cells, we identified plg7, encoding phospholipase A2, as a multicopy suppressor that restored the growth defect by removing RA from phospholipids and induced secretion of a part of the released free RA into culture media. In this study, we extended our analysis and examined the effect of triglyceride (TG) lipase overexpression on the tolerance to RA toxicity and RA productivity. S. pombe has three TG lipase genes, ptl1, ptl2, and ptl3, which have high protein sequence similarities to each other and to Saccharomyces cerevisiae counterparts TGL3, TGL4, and TGL5, but only ptl2 overexpression suppressed the growth defect induced by RA production, and the culture grown at 20?°C secreted free RA into media like plg7 overexpression. Suppression by ptl2 was independent of plg7, and a large amount of free RA was accumulated in the cells concomitant with the decrease in RA moieties in phospholipids. Furthermore, the suppression by ptl2 was attenuated by bromoenol lactone (BEL), a phospholipase A2 specific inhibitor, suggesting that Ptl2p may have phospholipase activity. Simultaneous overexpression of ptl2 and plg7 in the FAH12 integrant increased secretion and intracellular accumulation of RA 1.2- and 1.3-fold, respectively, compared to those with single overexpression of plg7 on day 10 at 20?°C.