An Engineered Escherichia coli Strain with Synthetic Metabolism for In-cell Production of Translationally Active Methionine Derivatives.

An Engineered Escherichia coli Strain with Synthetic Metabolism for In-cell Production of Translationally Active Methionine Derivatives. Chembiochem. 2020 Jul 31;: Authors: Schipp CJ, Ma Y, Al-Shameri A, D'Alessio F, Neubauer P, Contestabile R, Budisa N, di Salvo ML Abstract In the last decades, it became clear that the canonical amino acid repertoire codified by the universal genetic code is not up to the needs of the emerging biotechnologies. For such reason, extensive genetic code re-engineering is essential to expand the scope of ribosomal protein translation, leading to reprogrammed microbial cells equipped with an alternative biochemical alphabet to be exploited as potentially factories for biotechnological purposes. The prerequisite for this to happen is a continuous intracellular supply of non-canonical amino acids from simple and cheap precursors via synthetic metabolism. We engineered an E. coli bacterial system that fulfills these requirements through the reconfiguration of the methionine biosynthetic pathway and the introduction of an exogenous direct transsulfuration pathway. Our metabolic scheme operates in vivo, rescuing intermediates from core cell metabolism, and combining them with small bio-orthogonal compounds. Our reprogrammed E. coli strain is capable of in-cell production of L-azidohomoalanine, which is directly incorporated into proteins in response to methionine codons. We thereby constructed a prototype suit...
Source: Chembiochem - Category: Biochemistry Authors: Tags: Chembiochem Source Type: research