UDのエンジニアが、不自然なアミノ酸を合成できるバクテリアを作る(UD engineers create bacteria that can synthesize an unnatural amino acid)


2023-05-22 デラウェア大学 (UD)



生きた細菌で合成ニトロ化タンパク質を分散生産するためのプラットフォーム A platform for distributed production of synthetic nitrated proteins in live bacteria

Neil D. Butler,Sabyasachi Sen,Lucas B. Brown,Minwei Lin & Aditya M. Kunjapur
Nature Chemical Biology  Published:15 May 2023

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The incorporation of the nonstandard amino acid para-nitro-L-phenylalanine (pN-Phe) within proteins has been used for diverse applications, including the termination of immune self-tolerance. However, the requirement for the provision of chemically synthesized pN-Phe to cells limits the contexts where this technology can be harnessed. Here we report the construction of a live bacterial producer of synthetic nitrated proteins by coupling metabolic engineering and genetic code expansion. We achieved the biosynthesis of pN-Phe in Escherichia coli by creating a pathway that features a previously uncharacterized nonheme diiron N-monooxygenase, which resulted in pN-Phe titers of 820 ± 130 µM after optimization. After we identified an orthogonal translation system that exhibited selectivity toward pN-Phe rather than a precursor metabolite, we constructed a single strain that incorporated biosynthesized pN-Phe within a specific site of a reporter protein. Overall, our study has created a foundational technology platform for distributed and autonomous production of nitrated proteins.