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A rapid and sensitive enzymatic assay for 2,3-butanediol
Gyu Bi Lee1,2, Yun Jae Kim1,2, Jae Kyu Lim1,2
11Korea Institute of Ocean Science and Technology, Busan, 49111 Republic of Korea.
3 Biotech
|April 19, 2019
Summary
A new enzymatic assay rapidly detects 2,3-butanediol (2,3-BDO) with high sensitivity. This method is 15 times more sensitive than HPLC, enabling efficient screening of 2,3-BDO-producing microbes.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Microbiology
Background:
- Accurate detection of 2,3-butanediol (2,3-BDO) is crucial for identifying microbial production.
- Existing methods like High-Performance Liquid Chromatography (HPLC) have limitations in sensitivity and throughput.
Observation:
- A novel enzymatic assay was developed utilizing Clostridium ljungdahlii 2,3-butanediol dehydrogenase (CL-Bdh).
- The assay measures NADP+ reduction to quantify 2,3-BDO concentration.
Findings:
- The enzymatic assay detects 2,3-BDO at concentrations as low as 0.01 mM.
- This represents a 15-fold increase in sensitivity compared to HPLC, which requires >0.15 mM.
- In high-throughput screening, the enzymatic assay identified 14 positive microbial samples, versus 8 by HPLC.
Implications:
- The developed enzymatic assay offers a rapid, sensitive, and effective method for detecting 2,3-BDO.
- It is suitable for high-throughput screening of 2,3-BDO-producing microorganisms in a microtiter plate format.
- This advancement can accelerate research and development in bio-based chemical production.
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