Department of Chemical and Environmental Engineering, University of California, Riverside, CA 92521.
Department of Bioengineering, Stanford University, Stanford, CA 94305.
Proc Natl Acad Sci U S A. 2018 Apr 24;115(17):E3922-E3931. doi: 10.1073/pnas.1721469115. Epub 2018 Apr 2.
Microbial biosynthesis of plant natural products from simple building blocks is a promising approach toward scalable production and modification of high-value compounds. The pathway for biosynthesis of noscapine, a potential anticancer compound, from canadine was recently elucidated as a 10-gene cluster from opium poppy. Here we demonstrate the de novo production of noscapine in , through the reconstruction of a biosynthetic pathway comprising over 30 enzymes from plants, bacteria, mammals, and yeast itself, including 7 plant endoplasmic reticulum (ER)-localized enzymes. Optimization directed to tuning expression of pathway enzymes, host endogenous metabolic pathways, and fermentation conditions led to an over 18,000-fold improvement from initial noscapine titers to ∼2.2 mg/L. By feeding modified tyrosine derivatives to the optimized noscapine-producing strain we further demonstrated microbial production of halogenated benzylisoquinoline alkaloids. This work highlights the potential for microbial biosynthetic platforms to support the synthesis of valuable and novel alkaloid compounds, which can advance alkaloid-based drug discovery and development.
从简单的构建块微生物生物合成植物天然产物是一种有前途的方法，可以大规模生产和修饰高价值化合物。最近，从罂粟中发现了一种潜在的抗癌化合物北美黄连碱的生物合成途径，是一个由 10 个基因簇组成的途径。在这里，我们通过从植物、细菌、哺乳动物和酵母本身中重建一个包含 30 多种酶的生物合成途径，展示了在 中从头合成北美黄连碱，其中包括 7 种植物内质网（ER）定位酶。针对途径酶、宿主内源性代谢途径和发酵条件的优化，使初始北美黄连碱产量提高了 18000 多倍，达到约 2.2mg/L。通过向优化后的北美黄连碱生产菌株中添加修饰的酪氨酸衍生物，我们进一步证明了微生物可以生产卤代苯并异喹啉生物碱。这项工作突出了微生物生物合成平台在支持有价值和新型生物碱化合物合成方面的潜力，这可以推进基于生物碱的药物发现和开发。
