Phenotypic suppression of a fructose-1,6-diphosphate aldolase mutation in Escherichia coli.
Strain NP 315 of Escherichia coli possesses a thermolabile fructose-1, 6-diphosphate (FDP) aldolase; its growth on carbohydrate substrates is inhibited probably as a consequence of the accumulation of high intracellular levels of FDP. Studies of one class of phenotypic revertants of strain NP 315 which have regained their ability to grow on C(6) substrates at 40 C showed that in these strains the buildup of the inhibitory FDP pool is prevented by additional mutations in enzymes catalyzing the conversion of the substrate offered in the medium to FDP. For example, mutations affecting 6-phosphogluconate dehydrogenase activity (gnd(-)) may be selected in great number without any mutagenesis and enrichment simply by isolating revertants of strain NP 315 able to grow on gluconate at 40 C. Similarly, an additional mutation in phosphoglucose isomerase (pgi(-)) restores the ability of these fda(-)gnd(-) strains to grow on glucose at 40 C. Glucose metabolism of these fda(-)gnd(-)pgi(-) strains was investigated. The enzymes of the Entner-Doudoroff pathway are induced to an appreciable extent upon growth of these mutants on glucose medium; further evidence for glucose degradation via this route (which normally is induced only in the presence of gluconate) was provided by following the fate of the C1 label of radioactive glucose in l-alanine. Predominant labeling of the carboxyl-carbon of l-alanine was observed, inciating a major contribution of the Entner-Doudoroff path to pyruvate formation from glucose. Chromatographic analysis of the intermediates of glucose metabolism showed further that glucose apparently is at least partly metabolized via a bypass consisting of the accumulation of extracellular gluconic acid which arises by dephosphorylation of 6-phosphogluconolactone and possibly of 6-phosphogluconate. This extracellular gluconate is then taken up and metabolized in the normal manner via the Entner-Doudoroff enzymes.
D000409:Alanine / Q000378:metabolism; D002247:Carbon Isotopes; D002474:Cell-Free System; D002854:Chromatography, Paper; D003470:Culture Media; D004794:Enzyme Repression; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D005634:Fructose-Bisphosphate Aldolase / Q000096:biosynthesis* / Q000378:metabolism; D005636:Fructosephosphates / Q000378:metabolism; D005942:Gluconates / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism; D006836:Hydro-Lyases / Q000378:metabolism; D009154:Mutation; D010641:Phenotype; D010734:Phosphogluconate Dehydrogenase / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism; D013237:Stereoisomerism; D013696:Temperature
D002247:Carbon Isotopes; D003470:Culture Media; D005636:Fructosephosphates; D005942:Gluconates; D005958:Glucosephosphates; D011773:Pyruvates; D010734:Phosphogluconate Dehydrogenase; D010770:Phosphotransferases; D005634:Fructose-Bisphosphate Aldolase; D006836:Hydro-Lyases; D005956:Glucose-6-Phosphate Isomerase; D005947:Glucose; D000409:Alanine