Hoesl Michael Georg, Oehm Stefan, Durkin Patrick, Darmon Elise, Peil Lauri, Aerni Hans-Rudolf, Rappsilber Juri, Rinehart Jesse, Leach David, Söll Dieter, Budisa Nediljko
Institut für Chemie, Technische Universität Berlin, Müller-Breslau-Strasse 10, 10623 Berlin (Germany).
Institute of Cell Biology, School of Biological Sciences, University of Edinburgh (UK).
Angew Chem Int Ed Engl. 2015 Aug 17;54(34):10030-4. doi: 10.1002/anie.201502868. Epub 2015 Jul 1.
We have changed the amino acid set of the genetic code of Escherichia coli by evolving cultures capable of growing on the synthetic noncanonical amino acid L-β-(thieno[3,2-b]pyrrolyl)alanine ([3,2]Tpa) as a sole surrogate for the canonical amino acid L-tryptophan (Trp). A long-term cultivation experiment in defined synthetic media resulted in the evolution of cells capable of surviving Trp→[3,2]Tpa substitutions in their proteomes in response to the 20,899 TGG codons of the E. coli W3110 genome. These evolved bacteria with new-to-nature amino acid composition showed robust growth in the complete absence of Trp. Our experimental results illustrate an approach for the evolution of synthetic cells with alternative biochemical building blocks.
我们通过培养能够在合成的非标准氨基酸L-β-（噻吩并[3,2-b]吡咯基）丙氨酸（[3,2]Tpa）上生长的大肠杆菌培养物，改变了其遗传密码的氨基酸集，以此作为标准氨基酸L-色氨酸（Trp）的唯一替代物。在确定的合成培养基中进行的长期培养实验导致细胞进化，这些细胞能够响应大肠杆菌W3110基因组的20,899个TGG密码子，在其蛋白质组中进行Trp→[3,2]Tpa替换后存活下来。这些具有全新氨基酸组成的进化细菌在完全没有Trp的情况下显示出强劲的生长。我们的实验结果说明了一种利用替代生化构件进化合成细胞的方法。
