Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
Mar Drugs. 2013 Dec 11;11(12):5008-23. doi: 10.3390/md11125008.
Studies of polyunsaturated fatty acid (PUFA) biosynthesis in microalgae are of great importance for many reasons, including the production of biofuel and variable omega 3-long chain PUFAs. The elucidation of the PUFA biosynthesis pathway is necessary for bioengineering to increase or decrease PUFA content in certain microalgae. In this study, we identified the PUFA synthesis pathway in the oleaginous marine diatom, Fistulifera sp. strain JPCC DA0580, a promising candidate for biodiesel production. The data revealed not only the presence of the desaturases and elongases involved in eicosapentaenoic acid (EPA) synthesis, but also the unexpected localization of ω3-desaturase expression in the chloroplast. This suggests that this microalga might perform the final step of EPA synthesis in the chloroplast and not in the endoplasmic reticulum (ER) like other diatoms. The detailed fatty acid profile suggests that the EPA was synthesized only through the ω6-pathway in this strain, which was also different from other diatoms. Finally, the transcriptome analysis demonstrated an overall down-regulation of desaturases and elongases over incubation time. These genetic features might explain the decrease of PUFA percentage over incubation time in this strain. The important insights into metabolite synthesis acquired here will be useful for future metabolic engineering to control PUFA content in this diatom.
研究多不饱和脂肪酸(PUFA)在微藻中的生物合成具有重要意义,原因包括生物燃料的生产和可变的 ω3-长链多不饱和脂肪酸。阐明 PUFA 生物合成途径对于生物工程来说是必要的,以增加或减少某些微藻中的 PUFA 含量。在这项研究中,我们鉴定了产油海洋硅藻 Fistulifera sp. strain JPCC DA0580 中的 PUFA 合成途径,该硅藻是生产生物柴油的有前途的候选物。这些数据不仅揭示了参与二十碳五烯酸(EPA)合成的去饱和酶和延伸酶的存在,还揭示了 ω3-去饱和酶在叶绿体中的意外定位。这表明这种微藻可能在叶绿体中而不是在其他硅藻的内质网(ER)中完成 EPA 合成的最后一步。详细的脂肪酸谱表明,该菌株中 EPA 仅通过 ω6-途径合成,这与其他硅藻也不同。最后,转录组分析表明,随着孵育时间的延长,去饱和酶和延伸酶的整体表达下调。这些遗传特征可能解释了该菌株中 PUFA 百分比随孵育时间的下降。这里获得的对代谢物合成的重要见解将有助于未来对该硅藻中 PUFA 含量的代谢工程控制。