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Published on: August 8, 2016
A double-enzyme-coupled assay for high-throughput screening of succinic acid-producing strains
1Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
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.
