|
Descripción
|
|
|---|---|
| Backgrounds 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 the enzyme nitrogenase generates a minimum of one molecule of H2 per turnover. We present a method for the high-throughput selection of nitrogenase variants with enhanced H2 production. A Rhodobacter capsulatus strain has been re-engineered to generate a fluorescent signal in response to nitrogenaseproduced H2. A combination of random mutagenesis and fluorescence-activated cell sorting (FACS) is then used to select H2-overproducing mutants of R. capsulatus. Objectives To develop optimized biocatalysts for hydrogen production and to design a biotechnological tool for the detection of H2-overproducing bacteria. Methods The biotechnological tool consists of a library of (105-106) random nitrogenase variants and a biological H2 sensor, which is based on the sensing hydrogenase of R. capsulatus and in the engineered state produces fluorescent signals. High-throughput selection was achieved by coupling this tool to FACS cytometry. Conclusions We have re-engineered the natural H2 sensing system of R. capsulatus and combined it with FACS for the high-throughput selection of nitrogenase variants with enhanced H2 production that might independently retain their N2 fixation activity. This technology possesses great potential to identify nitrogenase amino acid substitutions leading to H2-overproducing variants that could be mimicked in nitrogenases from other microorganisms, expanding the impact of the findings. This tool was also used to perform a genome-wide screening of mutations leading to enhanced H2 production in R. capsulatus. | |
|
Internacional
|
Si |
|
Nombre congreso
|
FEMS 2017. 7th Congress of European Microbiologists |
|
Tipo de participación
|
970 |
|
Lugar del congreso
|
Valencia, España |
|
Revisores
|
Si |
|
ISBN o ISSN
|
00000000 |
|
DOI
|
|
|
Fecha inicio congreso
|
09/07/2017 |
|
Fecha fin congreso
|
12/07/2017 |
|
Desde la página
|
95 |
|
Hasta la página
|
95 |
|
Título de las actas
|
HYDROGEN OVERPRODUCING NITROGENASES OBTAINED BY RANDOM MUTAGENESIS AND HIGH-THROUGHPUT SCREENING |