Konzock Oliver, Tous-Mohedano Marta, Cibin Irene, Chen Yun, Norbeck Joakim
Division of Systems and Synthetic Biology, Department of Life Sciences, Chalmers University of Technology, Göteborg, Sweden.
AMB Express. 2023 Aug 10;13(1):84. doi: 10.1186/s13568-023-01590-3.
Yarrowia lipolytica has been explored as a potential production host for flavonoid synthesis due to its high tolerance to aromatic acids and ability to supply malonyl-CoA. However, little is known about its ability to consume the precursors cinnamic and p-coumaric acid. In this study, we demonstrate that Y. lipolytica can consume these precursors through multiple pathways that are partially dependent on the cultivation medium. By monitoring the aromatic acid concentrations over time, we found that cinnamic acid is converted to p-coumaric acid. We identified potential proteins with a trans-cinnamate 4-monooxygenase activity in Y. lipolytica and constructed a collection of 15 knock-out strains to identify the genes responsible for the reaction. We identified YALI1_B28430g as the gene encoding for a protein that converts cinnamic acid to p-coumaric acid (designated as TCM1). By comparing different media compositions we found that complex media components (casamino acids and yeast extract) induce this pathway. Additionally, we discover the conversion of p-coumaric acid to 4-hydroxybenzoic acid. Our findings provide new insight into the metabolic capabilities of Y. lipolytica and hold great potential for the future development of improved strains for flavonoid production.
解脂耶氏酵母因其对芳香酸的高耐受性和提供丙二酰辅酶A的能力，已被探索作为类黄酮合成的潜在生产宿主。然而，对于其消耗肉桂酸和对香豆酸前体的能力知之甚少。在本研究中，我们证明解脂耶氏酵母可以通过多种部分依赖于培养基的途径消耗这些前体。通过监测芳香酸浓度随时间的变化，我们发现肉桂酸被转化为对香豆酸。我们在解脂耶氏酵母中鉴定了具有反式肉桂酸4-单加氧酶活性的潜在蛋白质，并构建了一组15个敲除菌株以鉴定负责该反应的基因。我们鉴定出YALI1_B28430g为编码将肉桂酸转化为对香豆酸的蛋白质的基因（命名为TCM1）。通过比较不同的培养基组成，我们发现复杂培养基成分（酪蛋白氨基酸和酵母提取物）诱导了该途径。此外，我们发现了对香豆酸向4-羟基苯甲酸的转化。我们的研究结果为解脂耶氏酵母的代谢能力提供了新的见解，并为未来开发改良的类黄酮生产菌株具有巨大潜力。
