Ribose utilization with an excess of mutarotase causes cell death due to accumulation of methylglyoxal.
Methylglyoxal (MG) is a highly reactive metabolic intermediate, presumably accumulated during uncontrolled carbohydrate metabolism. The major source of MG is dihydroxyacetone phosphate, which is catalyzed by MG synthase (the mgs product) in bacteria. We observed Escherichia coli cell death when the ribose transport system, consisting of the RbsDACBK proteins, was overproduced on multicopy plasmids. Almost 100% of cell death occurs a few hours after ribose addition (>10 mM), due to an accumulation of extracellular MG as detected by (1)H-nuclear magnetic resonance ((1)H-NMR). Under lethal conditions, the concentration of MG produced in the medium reached approximately 1 mM after 4 h of ribose addition as measured by high-performance liquid chromatography. An excess of the protein RbsD, recently characterized as a mutarotase that catalyzes the conversion between the beta-pyran and beta-furan forms of ribose, was critical in accumulating the lethal level of MG, which was also shown to require ribokinase (RbsK). The intracellular level of ribose 5-phosphate increased with the presence of the protein RbsD, as determined by (31)P-NMR. As expected, a mutation in the methylglyoxal synthase gene (mgs) abolished the production of MG. These results indicate that the enhanced ribose uptake and incorporation lead to an accumulation of MG, perhaps occurring via the pentose-phosphate pathway and via glycolysis with the intermediates fructose 6-phosphate and glyceraldehyde 3-phosphate. It was also demonstrated that a small amount of MG is synthesized by monoamine oxidase.
Department of Life Sciences, Korea Advanced Institute of Science and Technology, Yusong-Ku, Taejon 305-701, Republic of Korea.
Insook
I
Kim
Eunjung
E
Kim
Seokho
S
Yoo
Daesung
D
Shin
Bumchan
B
Min
Jeeyeon
J
Song
Chankyu
C
Park
D017209:Apoptosis; D002238:Carbohydrate Epimerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006019:Glycolysis; D010427:Pentose Phosphate Pathway; D011765:Pyruvaldehyde / Q000378:metabolism*; D012266:Ribose / Q000378:metabolism*; D015854:Up-Regulation
D029968:Escherichia coli Proteins; D012266:Ribose; D011765:Pyruvaldehyde; D002238:Carbohydrate Epimerases; C019646:aldose 1-epimerase