title: A double-enzyme-coupled assay for high-throughput screening of succinic acid-producing strains.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
authors: L
authors: L
authors: Sun
authors: H
authors: H
authors: Zhang
authors: H
authors: H
authors: Yuan
authors: R
authors: R
authors: Tu
authors: Q
authors: Q
authors: Wang
authors: Y
authors: Y
authors: Ma
mesh_terms: D019342:Acetic Acid / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology*; D057075:Enzyme Assays / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology; D005285:Fermentation; D005561:Formates / Q000378:metabolism; D057166:High-Throughput Screening Assays; D009195:Peroxidase / Q000378:metabolism; D013385:Succinate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D019802:Succinic Acid / Q000378:metabolism*
chemical_list: D005561:Formates; C030544:formic acid; D019802:Succinic Acid; D009195:Peroxidase; D013385:Succinate Dehydrogenase; D019342:Acetic Acid