Department of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, 422-8526, Japan.
Chembiochem. 2022 Feb 16;23(4):e202100645. doi: 10.1002/cbic.202100645. Epub 2021 Dec 29.
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 in Aspergillus fumigatus to examine fungal metabolite production by HeLa cells that express fma-P450 heterologously. Here we show that HeLa cells harboring fma-P450 can 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.
生物合成基因不仅负责生物活性物质的形成，也适合于包括基因治疗在内的其他应用。为了测试在提供异源生物合成基因的情况下，人类细胞是否可以原位产生抗生素，我们专注于细胞色素 P450，这是一类在赋予天然产物前体生物活性方面很重要的酶。我们选择了在烟曲霉的 fumagillin 抗菌生物合成中起核心作用的 Fma-P450，以检查表达 Fma-P450 异源基因的 HeLa 细胞产生真菌代谢物的情况。在这里，我们表明，当用非毒性前体 β-trans-bergamotene 补充时，携带 Fma-P450 的 HeLa 细胞可以生物合成 5-羟-β-反式-bergamotene 和细胞毒性的 5-epi-去甲氧基 fumagillol。虽然产生的水平不足以导致细胞死亡，但我们证明通过引入异源生物合成基因来编程人类细胞自动生成抗生素是可行的。
