Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1186/s13068-025-02634-5
Lizenz creative commons licence
Titel (primär) Co-expression of auxiliary genes enhances the activity of a heterologous O2-tolerant hydrogenase in the cyanobacterium Synechocystis sp. PCC 6803
Autor Lupacchini, S.; Stauder, R.; Opel, F.; Klähn, S.; Schmid, A.; Bühler, B.; Toepel, J.
Quelle Biotechnology for Biofuels and Bioproducts
Erscheinungsjahr 2025
Department SOMA; MIBITECH
Band/Volume 18
Seite von art. 41
Sprache englisch
Topic T7 Bioeconomy
Supplements https://ndownloader.figstatic.com/files/53295503
Keywords Oxygen-tolerant hydrogenase; Oxygenic photosynthesis; Cyanobacteria; Hydrogenase maturation
Abstract Cyanobacteria bear great biotechnological potential as photosynthetic cell factories. In particular, hydrogenases are promising with respect to light-driven H2 production as well as H2-driven redox biocatalysis. Their utilization relies on effective strain design as well as a balanced synthesis and maturation of heterologous enzymes. In a previous study, the soluble O2-tolerant hydrogenase complex from Cupriavidus necator (CnSH) could be introduced into the model cyanobacterium Synechocystis sp. PCC 6803. Due to its O2-tolerance, it was indeed active under photoautotrophic growth conditions. However, the specific activity was rather low indicating that further engineering is required, for which we followed a two-step approach. First, we optimized the CnSH multigene expression in Synechocystis by applying different regulatory elements. Although corresponding protein levels and specific CnSH activity increased, the apparent rise in enzyme levels did not fully translate into activity increase. Second, the entire set of hyp genes, encoding CnSH maturases, was co-expressed in Synechocystis to investigate, if CnSH maturation was limiting. Indeed, the native CnSH maturation apparatus promoted functional CnSH synthesis, enabling a threefold higher H2 oxidation activity compared to the parental strain. Our results suggest that a fine balance between heterologous hydrogenase and maturase expression is required to ensure high specific activity over an extended time period
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=30608
Lupacchini, S., Stauder, R., Opel, F., Klähn, S., Schmid, A., Bühler, B., Toepel, J. (2025):
Co-expression of auxiliary genes enhances the activity of a heterologous O2-tolerant hydrogenase in the cyanobacterium Synechocystis sp. PCC 6803
Biotechnol. Biofuels Bioprod. 18 , art. 41 10.1186/s13068-025-02634-5