Metabolic Pathway Coupled with Fermentation Process Optimization for High-Level Production of Retinol in Yarrowia lipolytica,Journal of Agricultural and Food Chemistry

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Metabolic Pathway Coupled with Fermentation Process Optimization for High-Level Production of Retinol in Yarrowia lipolytica
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2024-04-02 , DOI: 10.1021/acs.jafc.4c00377
Xuefeng Ren _{1,

2,

3} , Mengsu Liu _{1,

2,

3} , Mingyu Yue _{1,

2,

3} , Weizhu Zeng _{1,

2,

3,

4} , Shenghu Zhou ₁ , Jingwen Zhou _{1,

2,

3,

4} , Sha Xu _{1,

3,

4}

Affiliation

Retinol is a lipid-soluble form of vitamin A that is crucial for human visual and immune functions. The production of retinol through microbial fermentation has been the focus of recent exploration. However, the obtained titer remains limited and the product is often a mixture of retinal, retinol, and retinoic acid, necessitating purification. To achieve efficient biosynthesis of retinol in Yarrowia lipolytica, we improved the metabolic flux of β-carotene to provide sufficient precursors for retinol in this study. Coupled with the optimization of the expression level of β-carotene 15,15′-dioxygenase, de novo production of retinol was achieved. Furthermore, Tween 80 was used as an extractant and butylated hydroxytoluene as an antioxidant to extract intracellular retinol and prevent retinol oxidation, respectively. This strategy significantly increased the level of retinol production. By optimizing the enzymes converting retinal to retinol, the proportion of extracellular retinol in the produced retinoids reached 100%, totaling 1042.3 mg/L. Finally, total retinol production reached 5.4 g/L through fed-batch fermentation in a 5 L bioreactor, comprising 4.2 g/L extracellular retinol and 1.2 g/L intracellular retinol. This achievement represents the highest reported titer so far and advances the industrial production of retinol.

中文翻译：

代谢途径与发酵工艺优化相结合，用于解脂耶氏酵母高水平生产视黄醇

视黄醇是维生素 A 的脂溶性形式，对人类视觉和免疫功能至关重要。通过微生物发酵生产视黄醇一直是近年来探索的焦点。然而，获得的效价仍然有限，并且产物通常是视黄醛、视黄醇和视黄酸的混合物，需要纯化。为了在解脂耶氏酵母中实现视黄醇的高效生物合成，我们在本研究中提高了β-胡萝卜素的代谢通量，为视黄醇提供足够的前体。结合β-胡萝卜素15,15'-双加氧酶表达水平的优化，实现了视黄醇的从头生产。此外，吐温80用作提取剂，丁基羟基甲苯作为抗氧化剂，分别提取细胞内视黄醇和防止视黄醇氧化。这一策略显着提高了视黄醇的产量水平。通过优化视黄醛转化为视黄醇的酶，生成的类视黄醇中细胞外视黄醇的比例达到100%，总计1042.3 mg/L。最终，通过在5L生物反应器中补料分批发酵，总视黄醇产量达到5.4g/L，其中包含4.2g/L胞外视黄醇和1.2g/L胞内视黄醇。这一成就代表了迄今为止报道的最高滴度，并推进了视黄醇的工业生产。

更新日期：2024-04-02

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