香叶基香叶醇是一种直链二萜化合物,也是合成多种药物和植物天然产物的重要前体。为实现香叶基香叶醇的高效生产,该研究首先强化甲羟戊酸途径基因表达构建了一株香叶基香叶醇产量为168.0 mg/L的菌株,其次通过引入外源香叶基香叶基焦磷酸合酶、下调角鲨烯合成竞争途径增加前体的供应,结合敲除不同的调控因子使香叶基香叶醇的产量达到678.2 mg/L,随后通过过表达磷酸戊糖途径基因和增加乙酰辅酶A的供应,为香叶基香叶醇的合成提供更多的辅因子和前体,增加融合基因拷贝数进一步提升香叶基香叶醇的产量至1 116.5 mg/L。最后在5 L生物反应器中补料分批发酵香叶基香叶醇产量达到11.3 g/L,为目前报道的最高水平。该研究为香叶基香叶醇的工业化生产提供了一定的参考,也为萜类天然产物的生物合成提供了有潜力的平台。
Geranylgeraniol (GGOH) is a straight-chain diterpenoid and a crucial precursor for the synthesis of various drugs and natural terpenoids.To achieve the efficient production of geranylgeraniol, this study first enhanced the gene expression of the mevalonate pathway and constructed a strain which can produce 168.0 mg/L of geranylgeraniol.Secondly, by introducing exogenous geranylgeranyl diphosphate synthase and downregulating the squalene synthesis competition pathway, the supply of precursors was increased.Combined with knocking out different regulatory factors, the yield of geranylgeraniol reached 678.2 mg/L.Subsequently, by overexpressing the phosphopentose pathway gene and increasing the supply of acetyl CoA, more cofactors and precursors were provided for the synthesis of geranylgeraniol.Increasing the copy number of fusion genes further improved the yield of geranylgeraniol to 1 116.5 mg/L.Finally, in a 5 L bioreactor, the production of geranylgeraniol by fed-batch fermentation reached 11.3 g/L, which is currently the highest reported level.This study provides a reference for the industrial production of geranylgeraniol and serves as a promising platform for the biosynthesis of natural terpenoids.
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