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通过代谢工程提高解脂耶氏酵母中的中链脂肪酸产量。

Increasing medium chain fatty acids production in Yarrowia lipolytica by metabolic engineering.

机构信息

LISBP, Université de Toulouse, CNRS, INRA, INSA, Toulouse, France.

INRA, UMR1318, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, Versailles, France.

出版信息

Microb Cell Fact. 2018 Sep 10;17(1):142. doi: 10.1186/s12934-018-0989-5.

Abstract

BACKGROUND

Oleaginous yeast Yarrowia lipolytica is an organism of choice for the development of biofuel and oleochemicals. It has become a chassis for metabolic engineering in order to produce targeted lipids. Understanding the function of key-enzymes involved in lipid metabolism is essential to design better routes for enhanced lipid production and for strains producing lipids of interest. Because medium chain fatty acids (MCFA) are valuable compounds for biokerosene production, we previously generated strains capable of producing MCFA up to 12% of total lipid content (Rigouin et al. in ACS Synth Biol 6:1870-1879, 2017). In order to improve accumulation and content of C14 fatty acid (FA), the elongation, degradation and accumulation of these MCFA in Yarrowia lipolytica were studied.

RESULTS

We brought evidence of the role of YALI0F0654 (YlELO1) protein in the elongation of exogenous or de novo synthesized C14 FA into C16 FA and C18 FA. YlELO1 deletion into a αFAS_IW expressing strain leads to the sole production of C14 FA. However, because this strain does not provide the FA essential for its growth, it requires being cultivated with essential fatty acids and C14 FA yield is limited. To promote MCFA accumulation in Y. lipolytica without compromising the growth, we overexpressed a plant diglyceride acyltransferase specific for MCFA and reached an accumulation of MCFA up to 45% of total lipid content.

CONCLUSION

We characterized the role of YlELO1 in Y. lipolytica by proving its involvement in Medium chain fatty acids elongation. We showed that MCFA content can be increased in Yarrowia lipolytica by promoting their accumulation into a stable storage form (triacylglycerides) to limit their elongation and their degradation.

摘要

背景

产油酵母解脂耶氏酵母是生物燃料和油脂化学品开发的首选生物。为了生产目标脂质,它已成为代谢工程的底盘。了解参与脂质代谢的关键酶的功能对于设计更好的途径来提高脂质产量和生产感兴趣的脂质的菌株是至关重要的。由于中链脂肪酸(MCFA)是生物航空煤油生产的有价值的化合物,我们之前生成了能够生产高达总脂质含量 12%的 MCFA 的菌株(Rigouin 等人,ACS Synth Biol 6:1870-1879,2017)。为了提高 C14 脂肪酸(FA)的积累和含量,研究了这些 MCFA 在解脂耶氏酵母中的伸长、降解和积累。

结果

我们证明了 YALI0F0654(YlELO1)蛋白在外源或从头合成的 C14 FA 伸长为 C16 FA 和 C18 FA 中的作用。在表达αFAS_IW 的菌株中删除 YlELO1 导致仅产生 C14 FA。然而,由于该菌株不能提供其生长所需的 FA,它需要用必需脂肪酸培养,并且 C14 FA 的产量是有限的。为了在不影响生长的情况下促进 MCFA 在解脂耶氏酵母中的积累,我们过表达了一种特异性针对 MCFA 的植物二酰基甘油酰基转移酶,达到了高达总脂质含量 45%的 MCFA 积累。

结论

我们通过证明 YlELO1 参与中链脂肪酸伸长来表征 Y. lipolytica 中的 YlELO1 作用。我们表明,通过促进它们积累到稳定的储存形式(三酰基甘油)中,可以增加解脂耶氏酵母中的 MCFA 含量,从而限制它们的伸长和降解。

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