Cloning and functional characterization of putative Escherichia coli ABC multidrug efflux transporter YddA.

Cloning and functional characterization of putative Escherichia coli ABC multidrug efflux transporter YddA. J Microbiol Biotechnol. 2020 Apr 29;: Authors: Feng Z, Liu D, Liu Z, Liang Y, Wang Y, Liu Q, Liu Z, Zang Z, Cui Y Abstract A putative multidrug efflux gene, yddA, was cloned from the Escherichia coli K-12 strain. A drug-sensitive strain of E. coli missing the main multidrug efflux pump AcrB was constructed as a host and the yddA gene was knocked out in wild type（WT）and drug-sensitive E. coli△acrB to study the yddA function. Sensitivity to different substrates of WT E.coli, E. coli△yddA, E. coli△acrB and E. coli△acrB△yddA strains was compared with minimal inhibitory concentration (MIC) assays and fluorescence tests. MIC assay and fluorescence test results showed that YddA protein was a multidrug efflux pump that exported multiple substrates. Three inhibitors, ortho-vanadate, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and reserpine, were used in fluorescence tests. Ortho-vanadate and reserpine significantly inhibited the efflux and increased accumulation of ethidium bromide and norfloxacin, while CCCP had no significant effect on YddA-regulated efflux. The results indicated that YddA relies on energy released from ATP hydrolysis to transfer the substrates and YddA is an ABC-type multidrug exporter. Functional study of unknown ATP-binding cassette (ABC) superfamily transporters in the model organism E. coli is ...
Source: Journal of Microbiology and Biotechnology - Category: Biotechnology Authors: Tags: J Microbiol Biotechnol Source Type: research