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Main Title: In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation
Author(s): Nojoumi, Saba
Ma, Ying
Schwagerus, Sergej
Hackenberger, Christian P. R.
Budisa, Nediljko
Type: Article
Language Code: en
Abstract: In this study, we report our initial results on in situ biosynthesis of S-allyl-l-homocysteine (Sahc) by simple metabolic conversion of allyl mercaptan in Escherichia coli, which served as the host organism endowed with a direct sulfhydration pathway. The intracellular synthesis we describe in this study is coupled with the direct incorporation of Sahc into proteins in response to methionine codons. Together with O-acetyl-homoserine, allyl mercaptan was added to the growth medium, followed by uptake and intracellular reaction to give Sahc. Our protocol efficiently combined the in vivo synthesis of Sahc via metabolic engineering with reprogrammed translation, without the need for a major change in the protein biosynthesis machinery. Although the system needs further optimisation to achieve greater intracellular Sahc production for complete protein labelling, we demonstrated its functional versatility for photo-induced thiol-ene coupling and the recently developed phosphonamidate conjugation reaction. Importantly, deprotection of Sahc leads to homocysteine-containing proteins—a potentially useful approach for the selective labelling of thiols with high relevance in various medical settings.
Issue Date: 9-May-2019
Date Available: 13-May-2019
DDC Class: 570 Biowissenschaften; Biologie
540 Chemie und zugeordnete Wissenschaften
Subject(s): biorthogonal conjugations
direct sulfhydration/transsulfuration pathway
methionine metabolism
non-canonical amino acids
reprogrammed translation
selective labelling
Sponsor/Funder: DFG, 414044773, Open Access Publizieren 2019 - 2020 / Technische Universität Berlin
DFG, 32049920, SFB 765: Multivalenz als chemisches Organisations- und Wirkprinzip: Neue Architekturen, Funktionen und Anwendungen
DFG, 390540038, EXC 2008: Vereinigung von Systemen in der Katalyse
Journal Title: International Journal of Molecular Sciences
Publisher: MDPI
Publisher Place: Basel
Volume: 20
Issue: 9
Article Number: 2299
Publisher DOI: 10.3390/ijms20092299
EISSN: 1422-0067
Appears in Collections:FG Biokatalyse » Publications

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