Combining Metabolic Engineering and Lipid Droplet Assembly to Achieve Campesterol Overproduction in Saccharomyces cerevisiae,Journal of Agricultural and Food Chemistry

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Combining Metabolic Engineering and Lipid Droplet Assembly to Achieve Campesterol Overproduction in Saccharomyces cerevisiae
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2024-02-22 , DOI: 10.1021/acs.jafc.3c09764
Zhijie Qin _{1,

2} , Yunliang Zhang _{1,

2} , Song Liu _{1,

2} , Weizhu Zeng _{1,

2} , Jingwen Zhou _{1,

2,

3} , Sha Xu ₁

Affiliation

Campesterol is a kind of important functional food additive. Therefore, stable and efficient campesterol biosynthesis is significant. Herein, we first knocked out the sterol 22-desaturase gene in Saccharomyces cerevisiae and expressed sterol Δ7-reductase from Pangasianodon hypophthalmus, obtaining a strain that produced 6.6 mg/L campesterol. Then, the modular expression of campesterol synthesis enzymes was performed, and a campesterol titer of 88.3 mg/L was achieved. Because campesterol is a lipid-soluble macromolecule, we promoted lipid droplet formation by exploring regulatory factors, and campesterol production was improved to 169.20 mg/L. Next, triacylglycerol lipase was used to achieve compartment campesterol synthesis. After enhancing the expression of sterol Δ7-reductase and screening cations, the campesterol titer reached 438.28 mg/L in a shake flask and 1.44 g/L in a 5 L bioreactor, which represents the highest campesterol titer reported to date. Metabolic regulation combined with lipid droplet engineering may be useful for the synthesis of other steroids as well.

中文翻译：

结合代谢工程和脂滴组装实现酿酒酵母中菜油甾醇的过量生产

菜油甾醇是一种重要的功能性食品添加剂。因此，稳定、高效的菜油甾醇生物合成具有重要意义。在此，我们首先敲除酿酒酵母中的甾醇22-去饱和酶基因，并表达下眼鲶中的甾醇Δ7-还原酶，获得产油菜甾醇6.6 mg/L的菌株。然后对菜油甾醇合成酶进行模块化表达，获得菜油甾醇效价88.3 mg/L。由于菜油甾醇是脂溶性大分子，我们通过探索调控因素促进脂滴形成，菜油甾醇产量提高至169.20 mg/L。接下来，使用三酰甘油脂肪酶来实现隔室菜油甾醇合成。经过增强甾醇Δ7-还原酶的表达和筛选阳离子后，菜油甾醇在摇瓶中达到438.28 mg/L，在5 L生物反应器中达到1.44 g/L，这是迄今为止报道的最高菜油甾醇效价。代谢调节与脂滴工程相结合也可能有助于其他类固醇的合成。

更新日期：2024-02-22

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