De novo biosynthesis of complex natural product sakuranetin using modular co-culture engineering
De novo biosynthesis of complex natural product sakuranetin using modular co-culture engineering
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DOI：
10.1007/s00253-020-10576-1
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发表时间：
2020-04-13
影响因子：
5
通讯作者：
Zhang, Haoran
中科院分区：
文献类型：
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作者：
Wang, Xiaonan;Li, Zhenghong;Zhang, Haoran
黄酮类化合物是植物和真菌天然产物中的一个大家族，其中许多化合物已被发现具有突出的生物活性。利用工程菌作为异源生物合成黄酮类化合物的替代宿主已被广泛研究。然而，目前的微生物生物合成策略主要依赖于使用一种微生物菌株来适应漫长而复杂的黄酮途径，这对生产优化提出了重大挑战。在这里，我们适应新兴的模块化共培养工程方法，合理设计，建立和优化大肠杆菌共培养从头生物合成黄酮类化合物樱花素从简单的碳底物葡萄糖。特别是两个E。使用大肠杆菌菌株以适应樱花素生物合成途径。上游菌株被工程化用于途径中间体对香豆酸生产，而下游菌株将对香豆酸转化为樱花醇。通过逐步优化共培养系统，我们能够在48 h内从5 g/L葡萄糖生产29.7 mg/L樱花醇，这显著高于传统的基于单一培养的方法的产量。在补料分批生物反应器中成功地放大了共培养生物合成，从而产生79.0 mg/L的樱花素。据我们所知，这是迄今为止报道的使用异源宿主E.杆菌这项工作的发现扩展了模块化共培养工程的适用性，以解决与复杂天然产物的异源生物合成相关的挑战。
Flavonoids are a large family of plant and fungal natural products, among which many have been found to possess outstanding biological activities. Utilization of engineered microbes as surrogate hosts for heterologous biosynthesis of flavonoids has been investigated extensively. However, current microbial biosynthesis strategies mostly rely on using one microbial strain to accommodate the long and complicated flavonoid pathways, which presents a major challenge for production optimization. Here, we adapt the emerging modular co-culture engineering approach to rationally design, establish and optimize an Escherichia coli co-culture for de novo biosynthesis of flavonoid sakuranetin from simple carbon substrate glucose. Specifically, two E. coli strains were employed to accommodate the sakuranetin biosynthesis pathway. The upstream strain was engineered for pathway intermediate p-coumaric acid production, whereas the downstream strain converted p-coumaric acid to sakuranetin. Through step-wise optimization of the co-culture system, we were able to produce 29.7 mg/L sakuranetin from 5 g/L glucose within 48 h, which is significantly higher than the production by the conventional monoculture-based approach. The co-culture biosynthesis was successfully scaled up in a fed-batch bioreactor, resulting in the production of 79.0 mg/L sakuranetin. To our knowledge, this is the highest bioproduction concentration reported so far for de novo sakuranetin biosynthesis using the heterologous host E. coli. The findings of this work expand the applicability of modular co-culture engineering for addressing the challenges associated with heterologous biosynthesis of complex natural products.