dc.contributor.author	Koch, Nikolaj G.
dc.contributor.author	Baumann, Tobias
dc.contributor.author	Nickling, Jessica H.
dc.contributor.author	Dziegielewski, Anna
dc.contributor.author	Budisa, Nediljko
dc.date.accessioned	2022-11-28T14:37:25Z
dc.date.available	2022-11-28T14:37:25Z
dc.date.issued	2022-10-24
dc.date.updated	2022-11-10T12:51:14Z
dc.description.abstract	Across scales, many biological phenomena, such as protein folding or bioadhesion and cohesion, rely on synergistic effects of different amino acid side chains at multiple positions in the protein sequence. These are often fine-tuned by post-translational modifications that introduce additional chemical properties. Several PTMs can now be genetically encoded and precisely installed at single and multiple sites by genetic code expansion. Protein nitration is a PTM of particular interest because it has been associated with several diseases. However, even when these nitro groups are directly incorporated into proteins, they are often physiologically reduced during or shortly after protein production. We have solved this problem by using an engineered Escherichia coli host strain. Six genes that are associated with nitroreductase activity were removed from the genome in a simple and robust manner. The result is a bacterial expression host that can stably produce proteins and peptides containing nitro groups, especially when these are amenable to modification. To demonstrate the applicability of this strain, we used this host for several applications. One of these was the multisite incorporation of a photocaged 3,4-dihydroxyphenylalanine derivative into Elastin-Like Polypeptides. For this non-canonical amino acid and several other photocaged ncAAs, the nitro group is critical for photocleavability. Accordingly, our approach also enhances the production of biomolecules containing photocaged tyrosine in the form of ortho-nitrobenzyl-tyrosine. We envision our engineered host as an efficient tool for the production of custom designed proteins, peptides or biomaterials for various applications ranging from research in cell biology to large-scale production in biotechnology.
dc.description.sponsorship	TU Berlin, Open-Access-Mittel – 2022	en
dc.identifier.eissn	2296-889X
dc.identifier.uri	https://depositonce.tu-berlin.de/handle/11303/17735
dc.identifier.uri	https://doi.org/10.14279/depositonce-16521
dc.language.iso	en
dc.rights.uri	http://creativecommons.org/licenses/by/4.0/
dc.subject.ddc	570 Biowissenschaften; Biologie	de
dc.subject.other	elastin-like polypeptides
dc.subject.other	MjTyrRS
dc.subject.other	ONBY
dc.subject.other	lambda red recombineering
dc.subject.other	Keio collection
dc.subject.other	nitroreduction
dc.subject.other	mussel-inspired adhesives
dc.subject.other	genetic code expansion
dc.title	Engineered bacterial host for genetic encoding of physiologically stable protein nitration
dc.type	Article
dc.type.version	publishedVersion
dcterms.bibliographicCitation.articlenumber	992748
dcterms.bibliographicCitation.doi	10.3389/fmolb.2022.992748
dcterms.bibliographicCitation.journaltitle	Frontiers in Molecular Biosciences
dcterms.bibliographicCitation.originalpublishername	Frontiers
dcterms.bibliographicCitation.originalpublisherplace	Lausanne
dcterms.bibliographicCitation.volume	9
tub.accessrights.dnb	free
tub.affiliation	Fak. 2 Mathematik und Naturwissenschaften::Inst. Chemie::FG Biokatalyse
tub.publisher.universityorinstitution	Technische Universität Berlin

Engineered bacterial host for genetic encoding of physiologically stable protein nitration

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