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Synthetic Biology – Metabolic Engineering pp 1–19Cite as

Orthogonal Protein Translation Using Pyrrolysyl-tRNA Synthetases for Single- and Multiple-Noncanonical Amino Acid Mutagenesis

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Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 162))

Abstract

To date, the two systems most extensively used for noncanonical amino acid (ncAA) incorporation via orthogonal translation are based on the Methanococcus jannaschii TyrRS/tRNA TyrCUA and the Methanosarcina barkeri/Methanosarcina mazei PylRS/tRNA PylCUA pairs. Here, we summarize the development and usage of the pyrrolysine-based system for orthogonal translation, a process that allows for the recombinant production of site-specifically labeled proteins and peptides. Via stop codon suppression in Escherichia coli and mammalian cells, genetically encoded biomolecules can be equipped with a great diversity of chemical functionalities including click chemistry handles, post-translational modifications, and photocaged sidechains.

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Authors and Affiliations

  1. Department of Chemistry, Berlin Institute of Technology/TU Berlin, Biocatalysis Group, Müller-Breslau-Straße 10 / Sekr. L 1, D-10623, Berlin, Germany

    Tobias Baumann, Matthias Exner & Nediljko Budisa

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  1. Tobias Baumann
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  2. Matthias Exner
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  3. Nediljko Budisa
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Correspondence to Nediljko Budisa .

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Editors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana, Illinois, USA

    Huimin Zhao

  2. Institut für Bioprozess- und Biosystemtechnik, Technische Universitðt Hamburg-Harburg, Hamburg, Germany

    An-Ping Zeng

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Baumann, T., Exner, M., Budisa, N. (2016). Orthogonal Protein Translation Using Pyrrolysyl-tRNA Synthetases for Single- and Multiple-Noncanonical Amino Acid Mutagenesis. In: Zhao, H., Zeng, AP. (eds) Synthetic Biology – Metabolic Engineering. Advances in Biochemical Engineering/Biotechnology, vol 162. Springer, Cham. https://doi.org/10.1007/10_2016_37

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