A modular pathway engineering strategy for the high-level production of β-ionone in Yarrowia lipolytica
A modular pathway engineering strategy for the high-level production of β-ionone in Yarrowia lipolytica
复制标题
在解脂耶氏酵母中高水平生产 β-紫罗兰酮的模块化途径工程策略
DOI：
10.1186/s12934-020-01309-0
复制
发表时间：
2020-02-27
影响因子：
6.4
通讯作者：
Yang, Xiaofeng
中科院分区：
文献类型：
--
作者：
Lu, Yanping;Yang, Qingyu;Yang, Xiaofeng
背景溶脂雅罗威酵母(Yarrowia liolytica，Yarrowia liolytica)是生产化学品和生物燃料的极具吸引力的细胞工厂。通过引入异源生物合成途径和修改内源途径，该细胞工厂已经研究了许多具有商业价值的天然产物的生产。然而，由于天然产物的合成代谢涉及漫长的途径和复杂的调控，将碳重新引导到目标化合物的产物中仍然是一项繁琐的工作，往往导致生产性能低下。结果将卷状毛霉的CarB和双功能的CARRP基因和矮牵牛的类胡萝卜素裂解双加氧酶1(CCD1)基因导入解脂Y.liolytica，使β-紫罗兰酮的产量低至3.5 mg/L。为了进一步提高β-紫罗兰酮的合成水平，我们采用模块化工程策略构建和优化了产β-紫罗兰酮的生物合成途径。在摇瓶发酵中，β-紫罗兰酮效价为358 mg/L，在补料分批发酵中，β-紫罗兰酮效价约为1g/L(接近基线菌株的280倍)。结论通过整合外源基因和天然基因的整合，构建了高效产β-紫罗兰酮的溶脂基因平台。对工程菌株进行了优化途径和发酵条件的模块化工程策略研究，获得了迄今为止细胞工厂报道的最高的β-紫罗兰酮效价。这一有效策略可用于促进解脂耶尔森菌中其他萜类化合物的生物合成。
Background The GRAS and oleaginous yeast Yarrowia lipolytica (Y. lipolytica) is an attractive cell factory for the production of chemicals and biofuels. The production of many natural products of commercial interest have been investigated in this cell factory by introducing heterologous biosynthetic pathways and by modifying the endogenous pathways. However, since natural products anabolism involves long pathways and complex regulation, re-channelling carbon into the product of target compounds is still a cumbersome work, and often resulting in low production performance. Results In this work, the carotenogenic genes contained carB and bi-functional carRP from Mucor circinelloides and carotenoid cleavage dioxygenase 1 (CCD1) from Petunia hybrida were introduced to Y. lipolytica and led to the low production of beta-ionone of 3.5 mg/L. To further improve the beta-ionone synthesis, we implemented a modular engineering strategy for the construction and optimization of a biosynthetic pathway for the overproduction of beta-ionone in Y. lipolytica. The strategy involved the enhancement of the cytosolic acetyl-CoA supply and the increase of MVA pathway flux, yielding a beta-ionone titer of 358 mg/L in shake-flask fermentation and approximately 1 g/L (similar to 280-fold higher than the baseline strain) in fed-batch fermentation. Conclusions An efficient beta-ionone producing GRAS Y. lipolytica platform was constructed by combining integrated overexpressed of heterologous and native genes. A modular engineering strategy involved the optimization pathway and fermentation condition was investigated in the engineered strain and the highest beta-ionone titer reported to date by a cell factory was achieved. This effective strategy can be adapted to enhance the biosynthesis of other terpenoids in Y. lipolytica.