Details zur Publikation |
| Kategorie | Datenpublikation |
| DOI | 10.5281/zenodo.15839120 |
| Lizenz |
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| Titel (primär) | Dataset for "Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803" [Data set] |
| Autor | Till, J.; Toepel, J.
; Sacco, D.; Bühler, B.; Appel, J.; Gutekunst, K. |
| Quelle | Zenodo |
| Erscheinungsjahr | 2025 |
| Department | MIBITECH; AME |
| Sprache | englisch |
| Topic | T7 Bioeconomy |
| Abstract | The development of hydrogenases capable of operating under oxygenic photosynthetic conditions remains a key challenge for sustainable biohydrogen production. In this study, we developed a series of chimeric NAD(+)-reducing [NiFe]-hydrogenases (SH) combining structural elements from the O-2-tolerant SH of Cupriavidus necator (CnSH) and the ferredoxin-interacting SH of Synechocystis sp. PCC6803 (SynSH). By engineering chimeric HoxU and HoxF subunits, we developed constructs-MixSH, Ch-HoxEF(Syn)+U-Cn, and Ch-HoxU(swapCTD)-that successfully couple the CnHoxYH hydrogenase module to the SynHoxEFU reductase module while retaining O-2 tolerance and enhancing interaction with reduced ferredoxin. The lithoautotrophic growth of C. necator confirmed the tolerance of these variants to O-2, while activity assays in Synechocystis demonstrated partial hydrogenase function, including H-2 consumption and fermentative H-2 production. Notably, Ch-HoxEF(Syn)+U-Cn retained ferredoxin interaction despite lacking the [4Fe4S](U4) cluster, showing [2Fe2S](F2) in HoxF as a functional ferredoxin-binding site. Moreover, we achieved the artificial integration of a [2Fe2S] cluster into CnHoxF and identified the CnHoxF N-terminal domain as structurally and functionally analogous to SynHoxE. Although electron transfer efficiency and activity in Synechocystis remained limited, this work validates the modular engineering of [NiFe]-hydrogenases, uniting O-2-tolerance with ferredoxin interaction and offering a foundational step toward photosynthesis-coupled H-2 production. |
| Verknüpfte UFZ-Textpublikationen | |
| dauerhafte UFZ-Verlinkung | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=31899 |
| Till, J., Toepel, J., Sacco, D., Bühler, B., Appel, J., Gutekunst, K. (2025): Dataset for "Engineering O2-tolerant chimeric hydrogenases optimized for ferredoxin coupling in Synechocystis sp. PCC 6803" [Data set] Zenodo 10.5281/zenodo.15839120 |
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