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敲除磷脂酶 A 基因提高产油酵母解脂耶氏酵母的脂积累。

Disrupting a phospholipase A gene increasing lipid accumulation in the oleaginous yeast Yarrowia lipolytica.

机构信息

Shandong Provincial Key Laboratory of Synthetic Biology, Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China.

University of Chinese Academy of Sciences, Beijing, China.

出版信息

J Appl Microbiol. 2021 Jan;130(1):100-108. doi: 10.1111/jam.14779. Epub 2020 Jul 28.

Abstract

AIMS

Phospholipase A (PLA ) is a diverse superfamily that hydrolyzes fatty acyl ester bonds at the sn-2 position of phospholipids. The correlation between phospholipid metabolism and the anabolism of neutral lipids remains unclear in yeasts. This study aims to explore the effects of PLA on lipid accumulation in the oleaginous yeast Yarrowia lipolytica.

METHODS AND RESULTS

This study identified an actively expressed phospholipase A gene (PLA2-3, YAIL0_E16060g) in Y. lipolytica by quantitative PCR analysis. The gene PLA2-3 was disrupted in the strain po1gΔKu70 by homologous recombination and in the strain po1g-G3 by a CRISPR-Cas9 system, which caused an increase in stress sensitivity while the cell growth was not altered under fermentative conditions. Lipid production was performed in both flasks and bioreactors. The results showed that the lipid titre and lipid content were improved over 25% and 8-30%, respectively, in PLA2-3 disrupted strains compared to the controls.

CONCLUSIONS

Disruption of the phospholipase PLA2-3 gene could effectively improve lipid production in Y. lipolytica.

SIGNIFICANCE AND IMPACT OF THE STUDY

This study presented a strategy on improving the lipid production of oleaginous yeasts and a similar strategy might be used in other oleaginous microbes.

摘要

目的

磷脂酶 A(PLA)是一个多样化的超家族,能够在磷脂的 sn-2 位置水解脂肪酸酯键。磷脂代谢与中性脂质合成之间的相关性在酵母中尚不清楚。本研究旨在探讨 PLA 对产油酵母解脂耶氏酵母脂质积累的影响。

方法和结果

通过定量 PCR 分析,本研究在解脂耶氏酵母中鉴定出一个表达活跃的磷脂酶 A 基因(PLA2-3,YAIL0_E16060g)。通过同源重组在 po1gΔKu70 菌株中以及通过 CRISPR-Cas9 系统在 po1g-G3 菌株中敲除 PLA2-3 基因,导致菌株在发酵条件下对压力的敏感性增加,但细胞生长没有改变。在摇瓶和生物反应器中进行了脂质生产。结果表明,与对照相比,PLA2-3 敲除菌株的脂质产量和脂质含量分别提高了 25%以上和 8-30%。

结论

敲除磷脂酶 PLA2-3 基因可有效提高解脂耶氏酵母的脂质生产能力。

研究的意义和影响

本研究提出了一种提高产油酵母脂质生产的策略,类似的策略可能用于其他产油微生物。

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