A double-enzyme-coupled assay for high-throughput screening of succinic acid-producing strains

Authors

  • L. Sun,

    1. Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    2. College of Life Sciences, University of Chinese Academy of Sciences, Beijing, China
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  • H. Zhang,

    1. Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
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  • H. Yuan,

    1. Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
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  • R. Tu,

    Corresponding author
    • Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
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  • Q. Wang,

    Corresponding author
    • Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
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  • Y. Ma

    1. Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
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Correspondence

Ran Tu and Qinhong Wang, Tianjin Institute of Industrial Biotechnology, CAS, 32 XiQiDao, Tianjin Airport Economic Area, Tianjin 300308, China.

E-mails: tu_r@tib.cas.cn and wang_qh@tib.cas.cn

Abstract

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.

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