Relationship between the Production of Spirosomes and Anaerobic Glycolysis Activity in Escherichia coli B

Abstract

The effects of culture conditions (aerobic or anaerobic) and glucose in the medium on the production of spirosomes in Escherichia coli B were studied by SDS-PAGE and electron microscopy. The M, of the spirosome of E. coli B was estimated to be 97000. Electron microscopy revealed that the amount of spirosomes derived from anaerobic cultures was about eightfold larger than that from aerobic cultures. In SDS-PAGE, the bands of spirosome protein derived from anaerobic cultures were more intense than those derived from aerobic cultures, either in peptone water or in Davis-Mingioli's minimal medium. With increased glucose concentration under aerobic conditions, the intensity of the band of spirosome protein was similar to that observed under anaerobic conditions in basal media. These results suggest that spirosome production by E. coli B is related to its anaerobic glycolysis activity.

Escherichia coli heat-shock proteins IbpA/B are involved in resistance to oxidative stress induced by copper

“…Purification of proteins for in vitro tests. After prolonged incubation of E. coli under oxygen depletion, AdhE forms large active oligomers which can be isolated from bacterial cell extracts by ultracentrifugation (Matayoshi et al, 1989;Kessler et al, 1992); therefore, we applied a one-step AdhE purification method. After testing different growth conditions, we found that almost homogeneous enzyme could be isolated from E. coli cells growing microaerobically at 25 uC, in M9 minimal medium with 0.2 % glucose.…”

DnaK dependence of mutant ethanol oxidoreductases evolved for aerobic function and protective role of the chaperone against protein oxidative damage in Escherichia coli

“…Residues 1-449 and 763-890 of AdhE are involved in its polymerization and may also be responsible for the propensity of the protein to aggregate amorphously. The biological significance of spirosomes, however, remains enigmatic (3,5,6).Despite the reversibility of the two NADH-coupled reactions catalyzed by AdhE, wild-type E. coli is unable to grow on ethanol as a sole source of carbon and energy, because the adhE gene is transcribed aerobically at lowered levels (7-9) and the half-life of AdhE activity is shortened during aerobic metabolism by metal-catalyzed oxidation (MCO). During this process, the hydroxyl radicals locally generated by the Fe 2ϩ bound to AdhE covalently attack the amino acid side chains near the active site (10, 11).…”

“…Residues 1-449 and 763-890 of AdhE are involved in its polymerization and may also be responsible for the propensity of the protein to aggregate amorphously. The biological significance of spirosomes, however, remains enigmatic (3,5,6).…”

Purification of spirosin from Lactobacillus reuteri DSM 20016 and partial characterization based on its amino acid sequences.

“…There have been many reports on morphological, biochemical and immunochemical studies of spirosome as quoted in our previous papers (Yamato et al, 1994(Yamato et al, , 1995. Though some functional aspects of spirosome have been described so far Matayoshi et al, 1989), its function is not yet well understood.…”