Mutants of Escherichia coli with a growth requirement for either lysine or pyridoxine.
Two distinct phenotypic classes of lysine requiring auxotrophs of Escherichia coli are described. Mutants of the LysA class produce little or no active diaminopimelic acid (DAP) decarboxylase and specifically require lysine for growth. Mutants of the LysB class produce a cryptic DAP decarboxylase which can be activated both in vivo and in vitro by higher than normal levels of its cofactor, pyridoxal 5'-phosphate. The LysB mutants have an alternate requirement for lysine or pyridoxine. Both LysA and LysB mutations map at 55 min, close to the thyA locus of E. coli. The association between pyridoxal phosphate and DAP decarboxylase appears to be much weaker in LysB mutants than in wild-type bacteria, and the mutant enzyme also sediments more slowly than wild-type enzyme in sucrose density gradients. The results suggest that the LysB mutations alter a specific region (or subunit) of the enzyme molecule which is needed to stabilize the binding of pyridoxal phosphate. These studies help to resolve certain contradictory observations on DAP decarboxylase reported earlier and may have relevance to pyridoxal phosphate enzymes in general. Prototrophic revertants of LysB mutants arise by second site mutations that result in increased availability of intracellular pyridoxal phosphate. These revertants appear to be derepressed for pyridoxine biosynthesis.
D000596:Amino Acids / Q000096:biosynthesis; D002262:Carboxy-Lyases / Q000096:biosynthesis; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002501:Centrifugation, Zonal; D002874:Chromosome Mapping; D003124:Colorimetry; D003470:Culture Media; D003956:Dialysis; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D005827:Genetics, Microbial; D008239:Lysine / Q000378:metabolism*; D009154:Mutation; D010867:Pimelic Acids / Q000096:biosynthesis; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D011736:Pyridoxine / Q000032:analysis / Q000378:metabolism; D012440:Saccharomyces / Q000378:metabolism; D013053:Spectrophotometry; D013395:Sucrose; D014161:Transduction, Genetic
D000596:Amino Acids; D003470:Culture Media; D010867:Pimelic Acids; D013395:Sucrose; D011732:Pyridoxal Phosphate; D002262:Carboxy-Lyases; D008239:Lysine; D011736:Pyridoxine