Toward Engineered Biosynthesis of Drugs in Human Cells
Toward Engineered Biosynthesis of Drugs in Human Cells
复制标题
人类细胞中药物的工程生物合成
DOI：
10.1002/cbic.202100645
复制
发表时间：
2021
期刊：
影响因子：
3.2
通讯作者：
Watanabe Kenji
中科院分区：
文献类型：
--
作者：
Matsuda Shinya;Tsunematsu Yuta;Matsushita Takuma;Ogata Yuji;Hachiya Shihomi;Kishimoto Shinji;Miyoshi Noriyuki;Watanabe Kenji
生物合成基因不仅负责生物活性物质的形成，而且还适用于包括基因治疗在内的其他应用。为了测试人类细胞在提供异源生物合成基因时原位生产抗生素的可行性，我们重点研究了细胞色素 P450，这是一类在赋予天然产物前体生物活性方面非常重要的酶。我们选择在烟曲霉的烟曲霉素抗菌生物合成中起核心作用的 Fma-P450 来检查异源表达 fma-P450 的 HeLa 细胞的真菌代谢产物的产生。在这里，我们表明，当补充无毒前体 β-反式香柠檬素时，携带 fma-P450 的 HeLa 细胞可以生物合成 5-羟基-β-反式香柠檬素和细胞毒性 5-表-去甲氧基烟曲霉醇。虽然生产水平不足以影响细胞死亡，但我们证明通过引入异源生物合成基因对人类细胞进行编程以自动生成抗生素是可行的。
Biosynthetic genes are not only responsible for the formation of bioactive substances but also suited for other applications including gene therapy. To test the feasibility of human cells producing antibiotics in situ when provided with a heterologous biosynthetic gene, we focused on cytochrome P450, the class of enzymes important in conferring bioactivity to natural product precursors. We selected Fma‐P450 that plays a central role in the fumagillin antimicrobial biosynthesis inAspergillus fumigatusto examine fungal metabolite production by HeLa cells that expressfma‐P450heterologously. Here we show that HeLa cells harboringfma‐P450can biosynthesize 5‐hydroxyl‐β‐trans‐bergamoten and cytotoxic 5‐epi‐demethoxyfumagillol when supplemented with the nontoxic precursor β‐trans‐bergamotene. While the production level was insufficient to effect cell death, we demonstrate that programming human cells to autogenerate antibiotics by introducing a heterologous biosynthetic gene is feasible.