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

Feng Zhenyue; Liu Defu; Liu Ziwen; Liang Yimin; Wang Yanhong; Liu Qingpeng; Liu Zhenhua; Zhongjing Zang; Cui Yu-Dong

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자료유형: 학술저널

저자정보: Feng Zhenyue (Heilongjiang Bayi Agricultural University) Liu Defu (Heilongjiang Bayi Agricultural University) Liu Ziwen (Heilongjiang Bayi Agricultural University) Liang Yimin (Heilongjiang Bayi Agricultural University) Wang Yanhong (Heilongjiang Bayi Agricultural University) Liu Qingpeng (Heilongjiang Bayi Agricultural University) Liu Zhenhua (Heilongjiang Bayi Agricultural University) Zhongjing Zang (Heilongjiang Bayi Agricultural University) Cui Yu-Dong (Heilongjiang Bayi Agricultural University)

저널정보: 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제30권 제7호

발행연도: 2020.1

수록면: 982 - 995 (14page)

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A putative multidrug efflux gene, yddA, was cloned from the Escherichia coli K-12 strain. A drugsensitive 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. coliΔacrB to study the yddA function. Sensitivity to different substrates of WT E.coli, E. coliΔyddA, E. coliΔacrB and E. coliΔacrBΔyddA 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 conducive to discovering new multidrug resista

#Multidrug efflux gene #yddA #Escherichia coli K-12 #ATP-binding cassette #multidrug exporter

참고문헌 (57)

참고문헌 신청

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Li XZ / 2015 / The challenge of efflux-mediated antibiotic resistance in gram-negative bacteria / Clin. Microbiol. Rev 28:337~418

Moussatova A / 2008 / ATP-binding cassette transporters in Escherichia coli / Biochim. Biophys. Acta 1778:1757~1771

Hrycyna CA / 1998 / Multidrug ABC transporters from bacteria to man : an emerging hypothesis for the universality of molecular mechanism and function / Drug Resist. Updat 1:81~83

Singh H / 2016 / ATP-dependent substrate transport by the ABC transporter MsbA is proton-coupled / Nat. Commun 7:12387

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