A fluorometric assay for high-throughput phosphite quantitation in biological and environmental matrices.
Analyst
; 148(15): 3650-3658, 2023 Jul 26.
Article
in En
| MEDLINE
| ID: mdl-37424451
ABSTRACT
Phosphite, the anion of phosphorus acid, is an important metabolite in the global biogeochemical phosphorus cycle and a phosphorus species with unique agricultural properties. As such, methods for detecting phosphite quantitatively and selectively are critical to evidencing phosphorus redox chemistry. Here, we present a fluorescence-based assay for phosphite, utilizing the NAD+-dependent oxidation of phosphite by phosphite dehydrogenase and the subsequent reduction of resazurin to resorufin. With the application of a thermostable phosphite dehydrogenase, a medium-invariant analytical approach, and novel sample preparation methods, the assay is capable of rapid and accurate phosphite quantification with a 3 µM limit of detection in a wide array of biologically- and environmentally-relevant matrices, including bacterial and archaeal cell lysate, seawater, anaerobic digester sludge, and plant tissue. We demonstrate the utility of the assay by quantitating phosphite uptake in a model crop plant in the presence or absence of a phosphite-oxidising strain of Pseudomonas stutzeri as a soil additive, establishing this bacterium as an efficient phosphite converting biofertilizer.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Phosphites
Language:
En
Journal:
Analyst
Year:
2023
Document type:
Article
Affiliation country:
United States