Construction and use of a Cupriavidus necator H16 soluble hydrogenase promoter (P SH ) fusion to gfp (green fluorescent protein)

W2409946200 · 수집 2026-10-04 18:12:26

T10030 Electrocatalysts for Energy Conversion

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Hydrogenases are metalloenzymes that reversibly catalyse the oxidation or production of molecular hydrogen (H 2 ). Amongst a number of promising candidates for application in the oxidation of H 2 is a soluble [Ni–Fe] uptake hydrogenase (SH) produced by Cupriavidus necator H16. In the present study, molecular characterisation of the SH operon, responsible for functional SH synthesis, was investigated by developing a green fluorescent protein (GFP) reporter system to characterise P SH promoter activity using several gene cloning approaches. A P SH promoter-gfp fusion was successfully constructed and inducible GFP expression driven by the P SH promoter under de-repressing conditions in heterotrophic growth media was demonstrated in the recombinant C. necator H16 cells. Here we report the first successful fluorescent reporter system to study P SH promoter activity in C. necator H16. The fusion construct allowed for the design of a simple screening assay to evaluate P SH activity. Furthermore, the constructed reporter system can serve as a model to develop a rapid fluorescent based reporter for subsequent small-scale process optimisation experiments for SH expression.

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Construction and use of a <i>Cupriavidus necator</i> H16 soluble hydrogenase promoter (P <sub>SH</sub> ) fusion to <i>gfp</i> (green fluorescent protein)

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