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A new approach for the generation and high-throughput detection of hydrogen-overproducing bacteria

The great interest for hydrogen (H2) as energy vector underlies research programs aimed to improving its sustainable biological production. During the process of biological nitrogen fixation (N2 - 2NH3) the enzyme nitrogenase generates a minimum of one molecule of H2 per turnover. This work, funded by the European Research Council, reveals high potential for nitrogenase improvement as biocatalyst for H2 production.

This work shows the development of a biotechnological tool for the detection of H2-overproducing bacteria. This tool consists of a library of (105-106) random nitrogenase variants and a biological H2 sensor, which is based on the sensing hydrogenase of Rhodobacter capsulatus and produces fluorescent signals that are proportional to the amount of H2 produced by each variant. High-throughput selection by coupling this tool to FACS cytometry was achieved. The selection of a nitrogenase variant resulting in 1000% improvement in bacterial H2 production is shown as proof of concept.



Original Paper:

Barahona, E; Jimenez-Vicente, E; Rubio, LM. 2016. "Hydrogen overproducing nitrogenases obtained by random mutagenesis and high-throughput screening". Scientific Reports. DOI: 10.1038/srep38291".

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