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Journal of Bacteriology, August 2005, p. 5782-5789, Vol. 187, No. 16
0021-9193/05/$08.00+0     doi:10.1128/JB.187.16.5782-5789.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Conversion of Methylglyoxal to Acetol by Escherichia coli Aldo-Keto Reductases

Junsang Ko,1 Insook Kim,1 Seokho Yoo,1 Bumchan Min,2 Kyungmin Kim,1 and Chankyu Park1*

Department of Life Sciences, Korea Advanced Institute of Science and Technology, Yusong-Ku, Taejon 305-701, Republic of Korea,1 Analytical Science Center, Samyang Central R & D Institute, 63-2, Hwaam-dong, Yusong-Ku, Taejon 305-717, Republic of Korea2

Received 2 March 2005/ Accepted 1 June 2005

Methylglyoxal (MG) is a toxic metabolite known to accumulate in various cell types. We detected in vivo conversion of MG to acetol in MG-accumulating Escherichia coli cells by use of 1H nuclear magnetic resonance (1H-NMR) spectroscopy. A search for homologs of the mammalian aldo-keto reductases (AKRs), which are known to exhibit activity to MG, revealed nine open reading frames from the E. coli genome. Based on both sequence similarities and preliminary characterization with 1H-NMR for crude extracts of the corresponding mutant strains, we chose five genes, yafB, yqhE, yeaE, yghZ, and yajO, for further study. Quantitative assessment of the metabolites produced in vitro from the crude extracts of these mutants and biochemical study with purified AKRs indicated that the yafB, yqhE, yeaE, and yghZ genes are involved in the conversion of MG to acetol in the presence of NADPH. When we assessed their in vivo catalytic activities by creating double mutants, all of these genes except for yqhE exhibited further sensitivities to MG in a glyoxalase-deficient strain. The results imply that the glutathione-independent detoxification of MG can occur through multiple pathways, consisting of yafB, yqhE, yeaE, and yghZ genes, leading to the generation of acetol.


* Corresponding author. Mailing address: Department of Life Sciences, Korea Advanced Institute of Science and Technology, Yusong-Ku, Taejon 305-701, Republic of Korea. Phone: 82-42-869-2629. Fax: 82-42-869-2610. E-mail: ckpark{at}kaist.ac.kr.


Journal of Bacteriology, August 2005, p. 5782-5789, Vol. 187, No. 16
0021-9193/05/$08.00+0     doi:10.1128/JB.187.16.5782-5789.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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