Aims
To develop a microplate‐based high‐throughput assay for the detection of succinic acid, which is widely used in many fields of industry. The assay can also be applied to screen high succinic acid‐producing strains.
Methods and Results
Here, we developed a double‐enzyme‐based succinic acid detection assay using fumarate reductase and peroxidase (named FRDP assay) to screen and isolate high succinic acid‐producing bacteria. The assay was performed in 96‐well microplate in a high‐throughput and had a well linear detection range of succinic acid from 0·05 mmol l−1 to 0·2 mmol l−1. Interference analysis results showed the complex media (LB),the by‐products (formic acid and acetic acid) of fermentation and a longer fermentation time interfere with the FRDP assay, while the mineral media (M9) and short fermentation time (<12 h) could minimize the interference.
Conclusions
The developed double‐enzyme‐coupled FRDP assay can be used in quick screening of succinic acid production.
Significance and Impact of the study
In this study, a novel high‐throughput screening method to detect succinic acid was developed. This screening method is useful to screen succinic acid‐producing bacteria and can be easily adjusted to screen different strains for the production of many other metabolites.