A whole-cell, high-throughput hydrogenase assay to identify factors that modulate [NiFe]-hydrogenase activity.

ABSTRACT: [NiFe]-hydrogenases have attracted attention as potential therapeutic targets or components of a hydrogen-based economy. [NiFe]-hydrogenase production is a complicated process that requires many associated accessory proteins that supply the requisite cofactors and substrates. Current methods for measuring hydrogenase activity have low throughput and often require specialized conditions and reagents. In this work, we developed a whole-cell high-throughput hydrogenase assay based on the colorimetric reduction of benzyl viologen to explore the biological networks of these enzymes in Escherichia coli We utilized this assay to screen the Keio collection, a set of nonlethal single-gene knockouts in E. coli BW25113. The results of this screen highlighted the assay's specificity and

SUBMITTER: Lacasse MJ 

PROVIDER: S-EPMC6802511 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature