Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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L-glutamine:D-fructose-6-phosphate Aminotransferase as a Key Protein Linked to Multidrug Resistance in E. coli KD43162

  • Lee, Sung-Eun (School of Applied Biosciences, Kyungpook National University) ;
  • Jung, Tae-Jeon (Department of Laboratory Medicine, Kosin University Gospel Hospital College of Medicine) ;
  • Park, Byeoung-Soo (Research Station, Nanotoxtech Inc.) ;
  • Kim, Byung-Woo (Department of Life Science and Biotechnology, Dongeui University) ;
  • Lee, Eun-Woo (Department of Life Science and Biotechnology, Dongeui University) ;
  • Kim, Hye Jin (Department of Dental Hygiene, Dongeui University) ;
  • Yum, Jong Hwa (Department of Clinical Laboratory Science, Dongeui University)
  • Received : 2015.05.21
  • Accepted : 2015.05.27
  • Published : 2015.09.30
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Abstract

A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

Keywords

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