Expression and Identification of the PadhE-1 from Thermoanaerobacter ethanolicus JW200 and Its Binding with adhE Promoter in vitro

Abstract:

Thermoanaerobacter ethanolicus produces ethanol from biomass derived substrates at temperatures above 70°C. AdhE, which exhibited bifunctional acetaldehyde/ethanol dehydrogenase activities, play a key role in producing ethanol. Proteins interacting with the transcriptional regulatory region (TRR) of adhE were for the first time isolated and identified by affinity chromatography. One of the proteins, PadhE-1, a homolog of LacI family, was cloned and expressed in Escherichia coli. Through electromobility shift assays, it was determined that the interaction between PadhE-1 and TRRadhE was specific. The research would be helpful to construct the bacteria with the extremely thermophilic and high ethanol-producing characteristics.

Key words:Thermoanaerobacter ethanolicus JW200, Regulator, Affinity chromatography, Electrophoretic mobility shift assay