Johannes Tyler W, DeSieno Matthew A, Griffin Benjamin M, Thomas Paul M, Kelleher Neil L, Metcalf William W, Zhao Huimin
Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Chem Biol. 2010 Jan 29;17(1):57-64. doi: 10.1016/j.chembiol.2009.12.009.
FR-900098 is a potent chemotherapeutic agent for the treatment of malaria. Here we report the heterologous production of this compound in Escherichia coli by reconstructing the entire biosynthetic pathway using a three-plasmid system. Based on this system, whole-cell feeding assays in combination with in vitro enzymatic activity assays reveal an unusual functional role of nucleotide conjugation and lead to the complete elucidation of the previously unassigned late biosynthetic steps. These studies also suggest a biosynthetic route to a second phosphonate antibiotic, FR-33289. A thorough understanding of the FR-900098 biosynthetic pathway now opens possibilities for metabolic engineering in E. coli to increase production of the antimalarial antibiotic and combinatorial biosynthesis to generate novel derivatives of FR-900098.
FR-900098是一种用于治疗疟疾的强效化疗药物。在此，我们报告了通过使用三质粒系统重建整个生物合成途径，在大肠杆菌中异源生产该化合物的情况。基于该系统，全细胞补料试验与体外酶活性试验相结合，揭示了核苷酸共轭的一种不寻常的功能作用，并导致了对先前未确定的后期生物合成步骤的完整阐明。这些研究还提出了第二种膦酸酯抗生素FR-33289的生物合成途径。对FR-900098生物合成途径的透彻理解，为在大肠杆菌中进行代谢工程以提高抗疟抗生素的产量以及进行组合生物合成以生成FR-900098的新型衍生物开辟了可能性。
