Department of Biosystems Science and Engineering, ETH Zürich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
Metab Eng. 2012 Jul;14(4):325-35. doi: 10.1016/j.ymben.2012.04.002. Epub 2012 Apr 21.
The biosynthesis of non-ribosomal peptides, many of which have pharmaceutical activities, is an evolutionary privilege of microorganisms. Capitalizing on the universal set of the Streptomyces lavendulae non-ribosomal peptide synthase BpsA and the Streptomyces verticillus 4'-phosphopantetheinyl transferase Svp, we have engineered Escherichia coli as well as mammalian cells, including human stem cells, to produce the blue 3,3'-bipyridyl pigment keto-indigoidine and the reduced colorless but fluorescent leuco-isoform. Detailed characterization of a tailored substrate-free chromogenic assay and FACS analysis showed that indigoidine's blue color and fluorescence could be reliably profiled in bacteria and mammalian cells using standard multiwell-compatible detection equipment. Besides serving as an inexpensive, reliable, versatile and easy-to-assay cross-kingdom reporter system, the potential of having mammalian cells produce non-ribosomal peptides, preferably ones with biopharmaceutical activities, may provide novel treatment opportunities in future gene- and cell-based therapies.
非核糖体肽的生物合成，其中许多具有药物活性，是微生物的一种进化特权。利用普遍存在的链霉菌 lavendulae 非核糖体肽合酶 BpsA 和链霉菌 verticillus 4'-磷酸泛酰巯基乙胺转移酶 Svp，我们已经成功地将大肠杆菌和哺乳动物细胞（包括人类干细胞）工程化，以生产蓝色 3,3'-联吡啶色素酮吲哚和还原的无色但荧光的白型。详细的无定制底物显色测定法和流式细胞术分析表明，使用标准的多孔兼容检测设备，可以在细菌和哺乳动物细胞中可靠地对吲哚的蓝色和荧光进行分析。除了作为一种廉价、可靠、多功能和易于检测的跨领域报告系统之外，使哺乳动物细胞产生非核糖体肽（最好是具有生物制药活性的肽）的潜力可能为未来的基因和细胞治疗提供新的治疗机会。
