Limonene, a valuable cyclic monoterpene, has been broadly studied in recent decades due to its wide application in the food, cosmetics and pharmaceutical industries. Engineering of the yeast Yarrowia lipolytica for fermentation of renewable biomass lignocellulosic hydrolysate may reduce the cost and improve the economics of bioconversion for the production of limonene. The aim of this study was to engineer Y. lipolytica to produce limonene from xylose and low-cost lignocellulosic feedstock. The heterologous genes XR and XDH and native gene XK encoding xylose assimilation enzymes, along with the heterologous genes tNDPS1 and tLS encoding orthogonal limonene biosynthetic enzymes, were introduced into the Po1f strain to facilitate xylose fermentation to limonene. The initially developed strain produced 0.44 mg/L of limonene in 72 h with 20 g/L of xylose. Overexpression of genes from the mevalonate pathway, including HMG1 and ERG12, significantly increased limonene production from xylose to ∼9.00 mg/L in 72 h. Furthermore, limonene production peaked at 20.57 mg/L with 50% hydrolysate after 72 h when detoxified lignocellulosic hydrolysate was used. This study is the first to report limonene production by yeast from lignocellulosic feedstock, and these results indicate the initial steps toward economical and sustainable production of isoprenoids from renewable biomass by engineered Y. lipolytica.

中文翻译：

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工程产油酵母解脂耶氏酵母以提高木糖和木质纤维素水解产物的柠檬烯产量。

柠檬烯是一种有价值的环状单萜，由于其在食品、化妆品和制药行业的广泛应用，近几十年来得到了广泛的研究。用于发酵可再生生物质木质纤维素水解物的酵母解脂耶氏酵母的工程化可以降低成本并提高用于生产柠檬烯的生物转化的经济性。本研究的目的是改造解脂耶氏酵母以从木糖和低成本木质纤维素原料中生产柠檬烯。编码木糖同化酶的异源基因XR和XDH和天然基因XK，以及异源基因tNDPS1和tLS将编码正交柠檬烯生物合成酶的编码引入 Po1f 菌株以促进木糖发酵成柠檬烯。最初开发的菌株在 72 小时内产生 0.44 mg/L 的柠檬烯和 20 g/L 的木糖。来自甲羟戊酸途径的基因（包括HMG1和ERG12 ）的过表达使木糖的柠檬烯产量在 72 小时内显着增加至约 9.00 mg/L。此外，当使用解毒的木质纤维素水解物时，72 小时后柠檬烯的产量峰值为 20.57 毫克/升，水解物为 50%。本研究首次报道了酵母从木质纤维素原料中生产柠檬烯，这些结果表明，通过工程化解脂耶氏酵母从可再生生物质中经济和可持续地生产类异戊二烯迈出了第一步.