The interaction of luciferase, flavin mono-nucleotide and long-chain aldehydes in the light reaction catalyzed by preparations of luminous bacteria (original) (raw)
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Probing of Potential Luminous Bacteria in Bay of Bengal and Its Enzyme Characterization
The present study dealt with the isolation, identification and enzyme characterization of potential luminous bacteria from water, sediment, squid, and cuttle fish samples of the Karaikal coast, Bay of Bengal, India during the study period September 2007 – August 2008. Bioluminescent strains were screened in SWC agar and identified using biochemical tests. As Shewanella henadai was found to be the most common and abundant species with maximum light emission [69,702,240 photons per second (pps)], the optimum ranges of various physicochemical parameters that enhance the luciferase activity in Shewanella hanedai were worked out. The maximum luciferase activity was observed at the temperature of 25o C (69,674,387 pps), pH of 8.0 (70,523,671 pps), salinity of 20 ppt (71,674,387 pps), incubation period of 16 h (69,895,714 pps), 4% peptone (70,895,152 pps) as nitrogen source, 0.9% glycerol (71,625,196 pps), and the ionic supplements of 0.3% CaCO3 (73,991,591 pps), 0.3% K2HPO4 (73,919,915 pps), and 0.2% MgSO4 (72,161,155 pps). Shewanella hanedai was cultured at optimum ranges for luciferase enzyme characterization. From the centrifuged supernatant, the proteins were precipitated with 60% ammonium sulfate, dialyzed, and purified using anion- exchange chromatography, and then luciferase was eluted with 500 mM phosphate of pH 7.0. The purified luciferase enzyme was subjected to SDS-PAGE and the molecular mass was determined as 78 kDa.
Luminous bacteria synthesize luciferase anaerobically
Archives of Microbiology, 1979
Four species of luminous bacteria, Photobacterium phosphoreum, P. leiognathi, P. fischeri and Beneckea harveyi (two strains of each), were shown to synthesize luciferase anaerobically. One of these, P. phosphoreum, produced as much luciferase anaerobically as it did aerobically, and all four species were found to grow almost equally rapidly under the two sets of conditions. Previous work with B. harveyi and P. fischeri had shown that aerobic luciferase synthesis can proceed only after an inhibitor in the complex medium has been removed and a species-specific autoinducer secreted. All strains tested also removed the inhibitor and secreted an autoinducer anaerobically. The small amount of luciferase produced anaerobically by some strains is thus apparently not due either to lack of removal of inhibitor or to insufficient production of autoinducer but may involve an oxygen-dependent control mechanism.
Microbiology, 1985
Two-dimensional diffusion gradients (of NaCl and H+ concentrations) were established in solid growth medium containing glycerol and yeast extract as major carbon sources. These were used to investigate conditions favourable for growth and bioluminescence of three species of luminous bacteria during incubation at different temperatures. Photobacterium leiognathi, Photobacterium phosphoreum and Vibriofischeri all grew over the entire salt range used [O-9-3 % (w/v) NaCl] and at pH values < 7 at the most favourable temperatures (20 "C, 20 "C and 15 "C respectively); upper and lower temperature limits for growth over a 72 h period were 30 and 10, 25 and 5, and 30 and 5 "C respectively. Bioluminescence was observed at all temperatures that supported growth; in P. leiognathi emission at 10 "C was hardly detectable even after 72 h, but at higher temperatures it occurred at all NaCl concentrations. Low pH values and high NaCl concentrations favoured luminescence in the other two organisms; after 48 h light emission decreased from the high pH and low NaCl regions of the gels. These results are discussed with reference to the symbiotic ( P . leiognathi, V.$sheri) or free-living ( P . phosphoreum) origins of the organisms studied. 0001-2622 0 1985 SGM Downloaded from www.microbiologyresearch.org by IP: 54.91.154.74 On: Sat, 29 Oct 2016 19:35:23 2866 P . W A T E R S AND D . LLOYD method developed by Wimpenny & Waters (1984) has been used in the present work, to examine the effects of pH values and NaCl concentrations on the growth and luminescence of three different luminous bacteria at different temperatures. As well as facilitating determination of optimal culture conditions, the method provides a uniquely convenient approach to a study of multifactorial interactions.
Characterization of the Phototrap in Photosynthetic Bacteria
Annals of the New York Academy of Sciences, 1975
8 Abbreviations: AUT particles, the phototrapcontaining fraction prepared by the combined alkaline, urea. Triton-X-100 method for membrane dissolution; AUT-e, electrophoretically-purified AUT particles. 297 298 rubrum by methods previously described. Bacteriopheophytin was prepared by adding HCl (final concentration = 0.25 N) t o a solution of bacteriochlorophyll in an acetone/methanol (7/2) at room temperature and recovering the desired product after 30 minutes by evaporating the solvent under reduced pressure. The bacteriopheophytin was dissolved in the acetone/methanol solvent (7/2) before adding it in an experiment. Chlorophyll a was purchased from the Sigma Chemical Company (St. Louis, Mo.). The preparation of Mn(II1) hematoporphyrin IX was previously reported.6 All glass apparatus for redox studies in the absence of air were patterned after those of Harbury7 and Loach. y 8 r 9 Electron paramagnetic resonance signals were measured with Varian E-3 and E-4 spectrometers (Varian Associates, Palo Alto, Calif.). Low temperature measurements were conducted with a liquid helium or liquid nitrogen accessory. The light used t o excite the sample was provided by a Xenon Corporation Model B Spectroscopic Micropulser employing a Suntron-6A lOOJ Flashtube (Xenon Corp., Medford, Mass.). The experimental arrangement was similar t o that previously described for measuring kinetic transients by epr.
Isolation of bioluminescent bacteria from marine organisms
2020
Bioluminescence is an emission of cold light by enzyme driven reaction within certain living organisms. The most abundant and widely distributed light emitting organisms are luminescent bacteria. Such organisms are either found as freeliving in the ocean or in symbiotic relationship with the marine host. To employ bioluminescence in environmental monitoring, isolation of bioluminescent bacteria from the two different marine samples (sea water sample and various organs of squid and fish) were collected from different sites of Veraval seashore and fish markets located nearby seashore respectively. The marine organisms used in the study were 20-25 days old. Cultivation media that were used for isolation were sea water agar (SWA), luminous agar (LA) and nutrient agar (NA); out of which SWA proved to be the most suitable medium for their growth and luminescence. No bioluminescent bacterium was found in water samples and total five bioluminescent bacteria were isolated from five different organs of fish and squid each. Out of these five isolates, two were selected based on their maximum light intensity. These two isolates, PBS1 and PBF1, were further characterized biochemically. PBS1 was able to utilize glucose, galactose, maltose and were tested positive for catalase and oxidase tests. Similar results were obtained in case of PBF1 except it was tested positive for urease urea but was unable to utilize glucose. Both isolates thrived at neutral pH and showed maximum bioluminescence. Effect of NaCl concentration on luminescence revealed that the two isolates were not able to grow in media devoid of NaCl and the luminescence was found to be maximum at 3 % (w/v) NaCl supplementation.