Research Group BIONUC (Biotechnology of Nutraceuticals and Bioactive Compounds), Departamento de Biología Funcional, Área de Microbiología, Universidad de Oviedo, Oviedo, Principality of Asturias, Spain.
Principality of Asturias, IUOPA (Instituto Universitario de Oncología del Principado de Asturias), Principality of Asturias, Spain.
Microb Cell Fact. 2023 Aug 29;22(1):167. doi: 10.1186/s12934-023-02172-5.
Naringenin is an industrially relevant compound due to its multiple pharmaceutical properties as well as its central role in flavonoid biosynthesis.
On our way to develop Streptomyces albidoflavus J1074 as a microbial cell factory for naringenin production, we have significantly increased the yields of this flavanone by combining various metabolic engineering strategies, fermentation strategies and genome editing approaches in a stepwise manner. Specifically, we have screened different cultivation media to identify the optimal production conditions and have investigated how the additive feeding of naringenin precursors influences the production. Furthermore, we have employed genome editing strategies to remove biosynthetic gene clusters (BGCs) associated with pathways that might compete with naringenin biosynthesis for malonyl-CoA precursors. Moreover, we have expressed MatBC, coding for a malonate transporter and an enzyme responsible for the conversion of malonate into malonyl-CoA, respectively, and have duplicated the naringenin BGC, further contributing to the production improvement. By combining all of these strategies, we were able to achieve a remarkable 375-fold increase (from 0.06 mg/L to 22.47 mg/L) in naringenin titers.
This work demonstrates the influence that fermentation conditions have over the final yield of a bioactive compound of interest and highlights various bottlenecks that affect production. Once such bottlenecks are identified, different strategies can be applied to overcome them, although the efficiencies of such strategies may vary and are difficult to predict.
由于柚皮素具有多种药物特性及其在类黄酮生物合成中的核心作用，因此它是一种具有工业相关性的化合物。
在将白色链霉菌 J1074 开发为生产柚皮素的微生物细胞工厂的过程中，我们通过逐步结合各种代谢工程策略、发酵策略和基因组编辑方法，显著提高了这种黄烷酮的产量。具体而言，我们筛选了不同的培养介质以确定最佳的生产条件，并研究了柚皮素前体的添加喂养如何影响生产。此外，我们还采用基因组编辑策略来去除与可能与柚皮素生物合成竞争丙二酰辅酶 A 前体的途径相关的生物合成基因簇（BGC）。此外，我们表达了 MatBC，分别编码丙二酸盐转运蛋白和负责将丙二酸盐转化为丙二酰辅酶 A 的酶，并复制了柚皮素 BGC，进一步促进了产量的提高。通过结合所有这些策略，我们能够使柚皮素的产量显著提高 375 倍（从 0.06mg/L 提高到 22.47mg/L）。
这项工作表明发酵条件对目标生物活性化合物最终产量的影响，并强调了影响生产的各种瓶颈。一旦确定了这些瓶颈，可以应用不同的策略来克服它们，尽管这些策略的效率可能会有所不同且难以预测。
