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      A nanoengineered tandem nitroreductase: designing a robust prodrug-activating nanoreactor†

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          Abstract

          Nitroreductases are important enzymes for a variety of applications, including cancer therapy and bioremediation. They often require encapsulation to improve stability and activity. We focus on genetically encoded encapsulation of nitroreductases within protein capsids, like encapsulins. Our study showcases the encapsulation of nitroreductase NfsB as functional dimers within encapsulins, which enhances protein activity and stability in diverse conditions. Mutations within the pore region are beneficial for activity of the encapsulated enzyme, potentially by increasing diffusion rates. Cryogenic electron microscopy reveals the overall architecture of the encapsulated dimeric NfsB within the nanoreactor environment and identifies multiple pore states in the shell. These findings highlight the potential of encapsulins as versatile tools for enhancing enzyme performance across various fields.

          Abstract

          Our study demonstrates that encapsulating tandem nitroreductase NfsB within encapsulins, along with pore mutations, enhances enzyme activity, with cryo-EM revealing its structural architecture and multiple pore states.

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          Most cited references3

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          Angew. Chem., Int. Ed.

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            Microbial Production of High-Value Products

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              Angew. Chem., Int. Ed.

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                Author and article information

                Journal
                RSC Chem Biol
                RSC Chem Biol
                CB
                RCBSAO
                RSC Chemical Biology
                RSC
                2633-0679
                4 November 2024
                2 January 2025
                4 November 2024
                : 6
                : 1
                : 21-35
                Affiliations
                [a ] Institute of Organic Chemistry, University of Freiburg 79104 Freiburg im Breisgau Germany claudia.jessen-trefzer@ 123456pharmazie.uni-freiburg.de
                [b ] School of Chemistry, University of Glasgow Glasgow G12 8QQ UK jesko.koehnke@ 123456lci.uni-hannover.de
                [c ] Institute of Pharmaceutical Science, Pharmaceutical Technology and Biopharmacy, University of Freiburg 79104 Freiburg im Breisgau Germany
                [d ] Department of Medicinal Chemistry, Martin-Luther University of Halle-Wittenberg 06120 Halle/Saale Germany
                [e ] Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Alexandria University Alexandria Egypt
                [f ] Institute for Biochemistry and Molecular Biology, ZBMZ, Faculty of Medicine, University of Freiburg 79104 Freiburg im Breisgau Germany
                [g ] Institute of Pharmaceutical Sciences, Chemical Epigenetics Group, University of Freiburg 79104 Freiburg im Breisgau Germany
                [h ] CIBSS – Centre for Integrative Biological Signalling Studies, University of Freiburg Schänzlestrasse 18 79104 Freiburg im Breisgau Germany
                [i ] Institute of Food Chemistry, Leibniz University Hannover 30167 Hannover Germany
                Author notes
                [‡]

                Equal contribution.

                Author information
                https://orcid.org/0000-0002-6361-7716
                https://orcid.org/0000-0002-8820-036X
                https://orcid.org/0000-0003-4216-8189
                Article
                d4cb00127c
                10.1039/d4cb00127c
                11532998
                39508026
                0a6a9647-453f-4b58-bca8-f84d3f80fe00
                This journal is © The Royal Society of Chemistry

                This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

                History
                : 15 June 2024
                : 3 October 2024
                Page count
                Pages: 15
                Funding
                Funded by: Deutsche Forschungsgemeinschaft, doi 10.13039/501100001659;
                Award ID: JE 782/3-1
                Award ID: JE 782/4-1
                Award ID: RTG 2202
                Award ID: 868/22 (469954457)
                Award ID: 868/23 (471614207)
                Award ID: SFB138
                Award ID: 380/109-1
                Award ID: INST 380/109-1
                Award ID: EXC-2189 (390939984)
                Award ID: Ju295/17-1 (490929945)
                Award ID: CRC992 (192904750)
                Funded by: European Research Council, doi 10.13039/501100000781;
                Award ID: ERC CoG 101002326
                Funded by: Ministry of Higher Education and Scientific Research, doi 10.13039/100017057;
                Award ID: Unassigned
                Funded by: China Scholarship Council, doi 10.13039/501100004543;
                Award ID: Unassigned
                Funded by: Medical Research Council, doi 10.13039/501100000265;
                Award ID: MC_PC_17135, MC_UU_00034/7, MR/X011879/1
                Funded by: Scottish Funding Council, doi 10.13039/501100000360;
                Award ID: H17007
                Categories
                Chemistry
                Custom metadata
                Paginated Article

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