Manipulation of the precursor supply for high-level production of longifolene by metabolically engineered Escherichia coli
Manipulation of the precursor supply for high-level production of longifolene by metabolically engineered Escherichia coli
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DOI：
10.1038/s41598-018-36495-w
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发表时间：
2019-01-14
影响因子：
4.6
通讯作者：
Liu, Huizhou
中科院分区：
文献类型：
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作者：
Cao, Yujin;Zhang, Rubing;Liu, Huizhou
长叶烯是一种天然存在的三环倍半萜，广泛应用于许多不同的领域。迄今为止，这种有价值的萜烯主要是从某些松脂的高沸点馏分中生产的。微生物生产可以是从天然植物来源提取的有前途的替代方案。在这里，我们提出的代谢工程策略，组装生物合成途径，长叶烯生产大肠杆菌。E.通过异源表达来自欧洲云杉的密码子优化的长叶烯合酶，使大肠杆菌产生长叶烯。通过不同的FPP焦磷酸酶增加法尼基焦磷酸（FPP）的代谢通量，使长叶烯产量增加1.8倍。通过引入外源甲羟戊酸途径实现了长叶烯生产的额外增强（高达2.64 mg/L）。在补料分批条件下，表现最好的菌株能够在5 L生物反应器中产生382 mg/L的长叶烯。这些结果证明了微生物发酵生产长叶烯的可行性，并可作为未来构建更强大菌株的基础。
Longifolene is a naturally occurring tricyclic sesquiterpene widely used in many different fields. Up to now, this valuable terpene was mainly manufactured from the high-boiling fraction of certain pine resins. Microbial production can be a promising alternative to the extraction from natural plant sources. Here, we present the metabolic engineering strategy to assemble biosynthetic pathway for longifolene production in Escherichia coli. E. coli was rendered to produce longifolene by heterologously expressing a codon optimized longifolene synthase from Picea abies. Augmentation of the metabolic flux to farnesyl pyrophosphate (FPP) by different FPP synthases conferred a 1.8-fold increase in longifolene production. An additional enhancement of longifolene production (up to 2.64 mg/L) was achieved by introducing an exogenous mevalonate pathway. Underfed-batch conditions, the best-performing strain was able to produce 382 mg/L of longifolene in a 5 L bioreactor. These results demonstrated the feasibility of producing longifolene by microbial fermentation and could serve as the basis for the construction of more robust strains in the future.