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d -2-Hydroxyglutarate ( d -2-HG) is a metabolite involved in many physiological metabolic processes. When  d -2-HG is aberrantly accumulated due to mutations in isocitrate dehydrogenase or  d -2-HG dehydrogenase, it functions in a pro-oncogenic manner and is thus considered a therapeutic target and biomarker in many cancers. In this study, DhdR from  Achromobacter denitrificans  NBRC 15125 is identified as an allosteric transcriptional factor that negatively regulates  d -2-HG dehydrogenase expression and responds to the presence of  d -2-HG. Based on the allosteric effect of DhdR, a  d -2-HG biosensor is developed by combining DhdR with amplified luminescent proximity homogeneous assay (AlphaScreen) technology. The biosensor is able to detect  d -2-HG in serum, urine, and cell culture medium with high specificity and sensitivity. Additionally, this biosensor is used to identify the role of  d -2-HG metabolism in lipopolysaccharide biosynthesis of  Pseudomonas aeruginosa , demonstrating its broad usages.

A d-2-hydroxyglutarate biosensor based on specific transcriptional regulator DhdR