- Glyoxal Detoxification in Escherichia coli K-12 by NADPH Dependent Aldo-keto Reductases
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Changhan Lee , Insook Kim , Chankyu Park
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J. Microbiol. 2013;51(4):527-530. Published online August 30, 2013
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DOI: https://doi.org/10.1007/s12275-013-3087-8
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Glyoxal (GO) and methylglyoxal (MG) are reactive carbonyl compounds that are accumulated in vivo through various pathways. They are presumably detoxified through multiple pathways including glutathione (GSH)-dependent/independent glyoxalase systems and NAD(P)H dependent reductases. Previously, we reported an involvement of aldo-ketoreductases (AKRs) in MG detoxification. Here, we investigated the role of various AKRs (YqhE, YafB, YghZ, YeaE, and YajO) in GO metabolism. Enzyme activities of the AKRs to GO were measured, and GO sensitivities of the corresponding mutants were compared. In addition, we examined inductions of the AKR genes by GO. The results indicate that AKRs efficiently detoxify GO, among which YafB, YghZ, and YeaE are major players.
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Citations
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- Mutations Upregulating the flhDC Operon of Escherichia coli K-12
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Changhan Lee , Chankyu Park
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J. Microbiol. 2013;51(1):140-144. Published online March 2, 2013
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DOI: https://doi.org/10.1007/s12275-013-2212-z
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755
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Bacterial motility is governed by the flhDC master operon that is under the control of factors like OmpR, LrhA, HdfR, and H-NS. Previously, derivatives of the wild-type MG1655 strain of E. coli K-12 with enhanced motility were found to contain insertion sequences (ISs) in the regulatory region of the flhDC operon. Here, we report that not only integrations of IS insertion sequences into the regulatory region of the flhDC operon, but also a missense mutation in the lrhA gene enhances motility by relieving transcriptional repression of the flhDC operon. Two novel IS insertions were found upstream of flhDC. So far, the relationships between the transacting factors and the cis-acting regulatory sequences associated with the flhDC operon have not been clearly established. In this study, it was found that effects of the cis- and trans-acting mutations were acting in parallel, suggesting their apparently independent regulation of flagellar expression.
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Citations
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BMC Microbiology.2013;[Epub] CrossRef
- Development of a Suicidal Vector-Cloning System Based on Butanal Susceptibility Due to an Expression of YqhD Aldehyde Reductase
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Changhan Lee , Chankyu Park
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J. Microbiol. 2012;50(2):249-255. Published online April 27, 2012
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DOI: https://doi.org/10.1007/s12275-012-1438-5
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666
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Abstract
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Previously, we observed butanal/propanal sensitivity of
Escherichia coli K-12 when cells overexpress YqhD protein,
a NADPH dependent aldehyde reductase, possibly due to
an accumulation of butanol/propanol in vivo as the reaction
products. Based on this finding, we developed a suicidal
vector-cloning system derived from pUC19, in which lacZ
was substituted with the yqhD gene. As a result, when foreign
DNA was inserted into its multiple cloning sites by disrupting
an expression of YqhD, the recombinants survived on
butanal/propanal containing plate, whereas cells containing
the YqhD vector died because of the alcohol production
by YqhD. The cloning efficiency, estimated based on colony
PCR and enzyme digestion, was achieved more than 90%
when the suicidal vector system was used. Moreover, the
plasmid vector itself was stably maintained in the cell, presumably
due to its ability to remove toxic aldehydes being
accumulated in E. coli cell by metabolic stress.
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Citations
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- Screening of Genes Related to Methylglyoxal Susceptibility
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Insook Kim , Joonho Kim , Bumchan Min , Changhan Lee , Chankyu Park
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J. Microbiol. 2007;45(4):339-343.
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DOI: https://doi.org/2563 [pii]
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Abstract
PDF
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Methylglyoxal (MG) is a reactive metabolite known to accumulate in certain physiological conditions. We attempted to isolate genes associated with this metabolite by genome-wide mutagenesis with TnphoA derivative. After screening on methylglyoxal-containing plate, we obtained insertions in three different genes, ydbD, yjjQ, and yqiI, which gave rise to reproducible MG-sensitive phenotypes in glyoxalase-deficient strain. In addition to its MG sensitivity, the insertion in yqiI exhibited an impaired motility resulting from a reduced flagellar expression.
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