Toward Engineered Biosynthesis of Drugs in Human Cells

Chembiochem. 2022 Feb 16;23(4):e202100645. doi: 10.1002/cbic.202100645. Epub 2021 Dec 29.

Abstract

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.

Keywords: Fumagillin; fungal natural products; heterologous biosynthesis; human cells; terpenes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antifungal Agents / chemistry
  • Antifungal Agents / metabolism
  • Antifungal Agents / pharmacology*
  • Aspergillus fumigatus / drug effects*
  • Cell Survival / drug effects
  • Cytochrome P-450 Enzyme System / metabolism*
  • Dose-Response Relationship, Drug
  • HeLa Cells
  • Humans
  • Microbial Sensitivity Tests
  • Molecular Structure
  • Sesquiterpenes / chemistry
  • Sesquiterpenes / metabolism
  • Sesquiterpenes / pharmacology*
  • Structure-Activity Relationship

Substances

  • 5-demethoxyfumagillol
  • Antifungal Agents
  • Sesquiterpenes
  • Cytochrome P-450 Enzyme System