It was found that both cosolvents at studied concentrations did not cause a significant change in conformation of bacterial luciferase. The molecular dynamics simulation was used to study the structure of bacterial luciferase surrounded by either water molecules solely or by mixture of water with various numbers of glycerol/sucrose molecules. Higher concentrations of glycerol, up to 30–40%, were found to reduce the efficiency of the reaction, while this effect was not observed in the media with sucrose. Increment of quantum yield in media with 10% of both osmolytes was shown. The effects of viscous media with glycerol and sucrose (10–40%) on the kinetics of the bacterial bioluminescent reaction have been investigated by stopped-flow technique. The overall light path appears to fall into the sensitized class of chemiluminescence mechanism, distinct from the dioxetanone types. Some rationalization of the mechanism has resulted from spatial structure determination, NMR of intermediates, and dynamic optical spectroscopy.
Besides these natural substrates, variable bioluminescence properties are found using other reactants such as flavin‐analogs or aldehydes, but results also depend on the luciferase type. A high energy species, the source of the exothermicity is unknown except that it is not a luciferin cyclic peroxide, a dioxetanone, as identified in the pathway of the firefly and the marine bioluminescence systems. Sufficient exothermicity equivalent to the energy of a blue photon, is available in the chemical oxidation of the aldehyde to the corresponding carboxylic acid, and a luciferase‐bound FMNHOOH is a key player. The luciferases all have close sequence homology and in vitro, a highly efficient light generation is obtained from these natural metabolites as substrates. There are many types of luciferase from species of bioluminescent bacteria originating from both marine and terrestrial habitats. The overall light path appears to fall into the sensitized class of chemilumines-cence mechanism, distinct from the dioxetanone types.Īfter more than one‐half century of investigations, the mechanism of bioluminescence from the FMNH2 assisted oxygen oxidation of an aliphatic aldehyde on bacterial luciferase, continues to resist elucidation. Some rationalization of the mechanism has resulted from spatial structure determination, NMR of inter-mediates and dynamic optical spectroscopy.
Besides these natural substrates, variable biolumi-nescence properties are found using other reactants such as flavin analogs or aldehydes, but results also depend on the luciferase type. A high energy species, the source of the exothermicity, is unknown except that it is not a luciferin cyclic peroxide, a dioxetanone, as identified in the pathway of the firefly and the marine bioluminescence systems. Sufficient exothermicity equivalent to the energy of a blue photon is available in the chemical oxidation of the alde-hyde to the corresponding carboxylic acid, and a luciferase-bound FMNH-OOH is a key player. The luciferases all have close sequence homology, and in vitro, a highly efficient light generation is obtained from these natural metabolites as sub-strates. There are many types of lucifer-ase from species of bioluminescent bacteria originating from both marine and terrestrial habitats. After more than one-half century of investigations, the mechanism of bioluminescence from the FMNH 2 assisted oxygen oxidation of an aliphatic aldehyde on bacterial luciferase continues to resist elucidation.
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