PMID:9489667
| Citation |
Tötemeyer, S, Booth, NA, Nichols, WW, Dunbar, B and Booth, IR (1998) From famine to feast: the role of methylglyoxal production in Escherichia coli. Mol. Microbiol. 27:553-62 |
|---|---|
| Abstract |
The enzyme methylglyoxal synthase (MGS) was partially purified from Escherichia coli extracts, and the amino-terminal sequence of candidate proteins was determined, based on the native protein being a tetramer of about 69 kDa. Database analysis identified an open reading frame in the E. coli genome, YccG, corresponding to a protein of 16.9 kDa. When amplified and expressed from a controlled promoter, it yielded extracts that contained high levels of MGS activity. MGS expressed from the trc promoter accumulated to approximately 20% of total cell protein, representing approximately 900-fold enhanced expression. This caused no detriment during growth on glucose, and the level of methylglyoxal (MG) in the medium rose to only 0.08 mM. High-level expression of MGS severely compromised growth on xylose, arabinose and glycerol. A mutant lacking MGS was constructed, and it grew normally on a range of carbon sources and on low-phosphate medium. However, the mutant failed to produce MG during growth on xylose in the presence of cAMP, and growth was inhibited. |
| Links | |
| Keywords |
Carbon-Oxygen Lyases/genetics; Carbon-Oxygen Lyases/isolation & purification; Carbon-Oxygen Lyases/metabolism; Escherichia coli/chemistry; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/growth & development; Gene Expression Regulation, Bacterial; Genes, Bacterial; Glucose/metabolism; Glycerol/metabolism; Molecular Sequence Data; Open Reading Frames; Phosphates/metabolism; Plasmids/genetics; Pyruvaldehyde/metabolism; Xylose/metabolism |
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