Rivollier Julie, Gosling Sandrine, Pezo Valérie, Heck Marie-Pierre, Marliere Philippe
Institute of Systems and Synthetic Biology, Genopole, 91000 Evry, France.
The European Syndicate of Synthetic Scientists ans Industrialists (TESSSI), 81 rue Réaumur, 75002 Paris, France.
ACS Omega. 2025 May 12;10(20):20171-20178. doi: 10.1021/acsomega.4c10463. eCollection 2025 May 27.
An experimental approach to implanting foreign chemical reactions into living cells is to select for the catalytic production of a vital building block such as an amino acid. Alkene metathesis is unknown in extant biochemistry, but it is emerging as a new type of reaction to be catalyzed by protein enzymes. Here, we show how the alkenic amino acid vinylglycine can be generated in a biocompatible reaction from 5-allyloxy-2-amino-pent-3-enoate (APE) by ring-closing metathesis catalyzed by a standard Hoveyda-Grubbs catalyst. The vinylglycine produced in situ is then used as a precursor of isoleucine and methionine, thus allowing the growth of strains of Escherichia coli requiring these essential amino acids. The robust nutritional screen we have developed paves the way for the directed evolution of genetically encoded metathesis enzymes and the chemical elaboration of metathesis coenzymes.
将外来化学反应植入活细胞的一种实验方法是选择催化产生诸如氨基酸等重要组成部分。烯烃复分解反应在现存生物化学中并不为人所知，但它正成为一种可由蛋白质酶催化的新型反应。在此，我们展示了如何通过标准霍维达-格鲁布斯催化剂催化的关环复分解反应，在生物相容性反应中由5-烯丙氧基-2-氨基-戊-3-烯酸酯（APE）生成烯基氨基酸乙烯基甘氨酸。原位产生的乙烯基甘氨酸随后用作异亮氨酸和蛋氨酸的前体，从而使需要这些必需氨基酸的大肠杆菌菌株得以生长。我们开发的强大营养筛选方法为基因编码复分解酶的定向进化和复分解辅酶的化学修饰铺平了道路。
