Escherichia coli ASKA clone library harboring tRNA-specific adenosine deaminase (tadA) reveals resistance towards xanthorrhizol

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Title
Escherichia coli ASKA clone library harboring tRNA-specific adenosine deaminase (tadA) reveals resistance towards xanthorrhizol
Author(s)
Yogiara; D Kim; J K Hwang; Jae Gu Pan
Bibliographic Citation
Molecules, vol. 20, no. 9, pp. 16290-16305
Publication Year
2015
Abstract
Xanthorrhizol is a potent antimicrobial compound isolated from the rhizome of Curcuma xanthorrhiza. However, the mechanism of xanthorrhizol action is unknown. To screen for probable target(s), we introduced the ASKA pooled-plasmid library into Escherichia coli W3110 imp4213 and enriched the library for resistant clones with increasing concentrations of xanthorrhizol. After three rounds of enrichment, we found nine genes that increased xanthorrhizol resistance. The resistant clones were able to grow in LB medium containing 256 μg/mL xanthorrhizol, representing a 16-fold increase in the minimum inhibitory concentration. Subsequent DNA sequence analysis revealed that overexpression of tadA, galU, fucU, ydeA, ydaC, soxS, nrdH, yiiD, and mltF genes conferred increased resistance towards xanthorrhizol. Among these nine genes, tadA is the only essential gene. tadA encodes a tRNA-specific adenosine deaminase. Overexpression of E. coli W3110 imp4213 (pCA24N-tadA) conferred resistance to xanthorrhizol up to 128 μg/mL. Moreover, overexpression of two tadA mutant enzymes (A143V and F149G) led to a twofold increase in the MIC. These results suggest that the targets of xanthorrhizol may include tadA, which has never before been explored as an antibiotic target.
Keyword
Food-grade antimicrobial compoundsTRNA-specific adenosine deaminaseXanthorrhizol
ISSN
1420-3049
Publisher
MDPI
DOI
http://dx.doi.org/10.3390/molecules200916290
Type
Article
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1. Journal Articles > Journal Articles
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