Bioluminescence marine bacteria

Figure 2 Bioluminescence marine bacteria Beneckea harveyi. (Reproduced with permission of Ul Kricka, Hospital of the University of Pennsylvania, Philadelphia, USA.)...

Miyamoto, C., Boylan, M., Graham, A., and Meighen, E. (1986). Cloning and expression of the genes from the bioluminescent system of marine bacteria. Method. Enzymol. 133 70-83. 

Although numerous luminous organisms are known, only a few of them has been studied and really exploited. Analytical applications of bioluminescence concern mainly the detection of ATP with the firefly luciferase and of NADH with some marine bacteria systems. Luciferase from the North American firefly, i.e., Photinus pyralis, has been extensively studied10-12 and afterwards, attention has been paid to the luciferase from Luciola mingrelica, i.e., the North Caucasus firefly13 15. 

The bacterial bioluminescent reaction is also catalyzed by a luciferase (EC 1.14.14.3) isolated from marine bacteria. The four most studied types are Vibrio harveyi, Vibrio fischeri, Photobacterium phosphoreum and Photobacterium leiognathi18, 19. In these different luminescent bacteria the... 

The biological classification schemes for bacteria and archaea are still being developed because of the rapid pace of new discoveries in genomics. The two most important phyla of marine bacteria are the cyanobacteria, which are photosynthetic, and the proteobacteria. The latter include some photosynthetic species, such as the purple photosynthetic bacteria and N2 fixers. Other members of this diverse phylum are the methanotrophs, nitrifiers, hydrogen, sulfur and iron oxidizers, sulfete and sulfur reducers, and various bioluminescent species. 

Production of light by certain marine bacteria. The general consensus is that light is produced when bacterial luciferase catalyzes the bioluminescent oxidation of FMNH2 and a long chain aldehyde by molecular oxygen. Volume 1(1,2). 

Fourth, no ideal way to calibrate the photometers has been found. Seliger et al. (25) calibrated their pump-through bathyphotometer with dilute solutions of luminous marine bacteria. They assumed that the average spatial distribution of stimulated bioluminescence inside the impeller housing of their bathyphotometer was the same as that of the continuously luminous bacteria. They noted... 

Most of bacterial biosensors are based on the operon luxCDABE that codes for the bacterial luciferase founded in the marine bacteria V. fischeri and V. harveyi, and for an essential aldehyde substrate that would otherwise have to be supplied exogenously. The cluster luxAB cassette codes for the luciferase whereas luxCDE encodes a fatty acid reductase complex. The latter enzymes are responsible for the synthesis of the long-chain aldehyde that is required as substrate in the bioluminescence reaction (Meighen and Dunlap, 1993 Hakkila et al., 2002). Luciferase catalyses the oxidation reaction of flavin mononucleotide (FMNH2). A long-chain (7 to 16 carbons) aldehyde is reduced in presence of oxygen by the aldehyde reductase. The outcome of the bioluminescent reaction can be expressed as follows ... 

Quantitative criteria of expression efficiency of bioluminescence were copy number of lux-operon (level of a replication) in a cell, duration of the latent period of induction of bioluminescence (level of a transcription), the maximal intensity of luminescence (level of a translation). The following quantitative criteria and coefficients are used fi - number of copies of /wx-operons in a cell (for natural marine bacteria - 1 operon/chromosome, for transgenic luminous bacteria the parameter varies depending on strain). Number of copies of plasmid was determined on electrophoregrams with program Scion Image f2 = 1/t - coefficient (s 1), which reflects influence of the latent period in dynamics of bioluminescence on expression... 

If luminescence is a result of a biochemical reaction, the principle is called bioluminescence. The most frequently used bioluminescence system is that of the firefly. The enzyme luciferase catalyses the oxidation of luciferin as a substrate in the presence of adenosine triphosphate (ATP) (Scheme 7). Another bioluminescence system makes use of a luciferase from certain marine bacteria. A long-chain aldehyde is oxidized in the presence of luciferase, an oxido-reductase and NAD/NADH. Recently, a photoprotein isolated from the bioluminescent jellyfish Aequorea victoria, has been found to be an efficient bioluminescence label for immunoassays. 

Standardised test methods, such as ISO 11348 [59], could be used for aqueous samples and elutriates. Light-emitting marine bacteria, such as Vibrio fischeri or Photobacterium sp., are used. A defined bacterial inoculum is added to the sample solutions and the change of bioluminescence intensity is measured over a period of 30 min. Ready to use test kits, e.g., LumisTox (Dr. Lange) or ToxAlert (Merck) are available and comply with all the requirements defined in the standard methods. 

Bioluminescence can be used for spedfic detection of separated bioactive compounds on layers (BioTLC) [46]. After development and drying the mobile phase by evaporation, the layer is coated with microorganisms by immersion of the plate. Single bioactive substances in multicomponent samples are located as zones of differing luminescence. The choice of the luminescent cells determines the specificity of detection. A specific example is the use of the marine bacterium Vibrio fischeri with the BioTLC format. The bioluminescence of the bacteria cells on the layer is reduced by toxic substances, which are detected as dark zones on a fluorescent background. BioTLC kits are available from ChromaDex, Inc. (Santa Ana, CA). 

The more than 6000 species show various mechanisms of defense against predators. In marine species, amphipod luminescence always seems to be a defensive response.88 In the freshwater species Hyalella azteca, a bioluminescence caused by bacteria was reported.89... 

Transgenic bacterial biosensors. Systems such as the Microtox assay detailed earlier use the marine species Vibrio fischeri as the sensor. Because it uses a marine bacterium, Microtox must be conducted in saline solution, which is ecologically irrelevant for most soils. Because no naturally luminescent soil bacteria are known that could be used as an alternative, one solution is to fuse the genes responsible for bioluminescence into soil-dwelling strains using recombinant technology (Paton et al., 1997). Two approaches can be used ... 

Luminous bacteria are bioluminescent microorganisms whose luciferase genes (lux), proteins and intact cells are widely used in applied research and commercial products. Acknowledging the commercial value of luminescent cells also in entertainment and education, we have conducted research on luminous bacteria from marine samples and have isolated Photobacterium phosphoreum (strain RL-1) from coastal marine sediment. In order to maximize the luminescence activity of RL-1, we examined a series of extracts prepared from dried marine foodstuff. Because chitinous compounds and some amino acids are known to be abundant in dried squid and shrimp, we also tested the effects of those compounds on the luminescence activity. Among the supplemental compounds tested, chitosan, cysteine, and aspartic acid were found to enhance the luminescence activity of RL-1. The present results indicate that some amino acids and chitinous compounds are effective supplements for further enhancing bacterial light production in an enriched medium (SWC ). 

The Beckman Microtox system was employed to assess the relative toxicity of pesticides and their hydrolysis products to bacteria. This system utilizes Photobacterium phosphoreum, a marine bioluminescent bacterium phylogenetically related to several genera of bacteria important in soil. The Microtox system measures the light emitted from P. phosphoreum that have been exposed to a chemical dissolved in the diluent. The details of theory and operation of Microtox analyzer and experimental conditions used have been described (26-28). 

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