Escherichia coli ASKA Clone Library Harboring tRNA-Specific Adenosine Deaminase (tadA) Reveals Resistance towards Xanthorrhizol

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...

Full description

Saved in:
Bibliographic Details
Published in:Molecules (Basel, Switzerland) Switzerland), 2015-09, Vol.20 (9), p.16290-16305
Main Authors: Yogiara, Kim, Dooil, Hwang, Jae-Kwan, Pan, Jae-Gu
Format: Article
Language:English
Subjects:
Adenosine
Adenosine Deaminase - genetics
Adenosine Deaminase - metabolism
Anti-Bacterial Agents - pharmacology
Antimicrobial agents
Bacteria
Biotechnology
Cloning
Curcuma
E coli
Escherichia coli
Escherichia coli - drug effects
Escherichia coli - genetics
food-grade antimicrobial compounds
Fucus
Genes
Genomics
Gram-negative bacteria
Gram-positive bacteria
Microbial Sensitivity Tests
Phenols - pharmacology
Plasmids
RNA, Transfer - genetics
Transfer RNA
tRNA-specific adenosine deaminase
xanthorrhizol
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1420-3049
1420-3049
DOI:10.3390/molecules200916290