Development of a Highly Sensitive, Fast and Efficient Screening Technique for the Detection of 2,3-Butanediol by Thin Layer Chromatography

Abstract

The interest in microbial production of 2,3-butanediol is based on the awareness that 2,3 butanediol is a promising bulk chemical due to its extensive industrial applications. Here, we report a novel method for the detection of 2,3-butanediol. The proposed method has a wide applicability to harness the commercial potential of microorganisms which produce 2,3-butanediol as the end product. Experimentally 32 bacterial strains were screened for 2,3-butanediol production. After 72 h the samples were spotted on thin-layer chromatography plates and ran in a solvent system comprising of hexane:ethyl-acetate:glacial acetic acid in the ratio of 70:30:1.5, followed by colour development using vanillin reagent. The appearance of a blue-colored spot of 2,3-butanediol with a retention factor (R f) of 0.68 corresponds to the R f value of the standard 2,3-butanediol which forms the basis for the selection of 2,3-butanediol producers. Apart from being a rapid detection system the proposed method is highly sensitive as it was able to detect a concentration as low as 1.0 mg/ml and its authenticity was reconfirmed by GC and GC-MS. Low cost of this method provides an effective support for 2,3-butanediol detection at any scale.

A rapid and sensitive enzymatic assay for 2,3-butanediol

“…Thus, GC and HPLC are not suitable for screening purposes. Recently, a method to detect 2,3-BDO using thin-layer chromatography (TLC) was suggested (Saran et al 2014), which is simple and highly sensitive but not suitable for the high-throughput screening system. In this report, we present a new enzymatic assay for highthroughput screening of 2,3-BDO, using 2,3-butanediol dehydrogenase from Clostridium ljungdahlii DSM 13528.…”