RESEARCH AREAS AND PRACTICAL APPLICATION
OF LUMINOUS BACTERIA
The research work in the collection done along the
preservation of viability and initial properties of the Collection gene pool;
of optimum methods of gene pool
physiological, biochemical, and morphological properties of collection strains;
of strains producing luciferases, oxidoreductases and other target products,
including, using of E.coli as heterologic owner of such products;
distribution of these microorganisms in waters of the World ocean and their
population heterogeneity and dissociation under various cultivation conditions;
structural organization and fatty acid composition of luminous bacteria.
luminescent system localization;
of the mechanism of toxicant action on the cell and the population as a whole;
activity and stability of enzymes of the luminescence reaction;
information network, computer databases of microorganisms;
of high-sensitive toxicity biotest for monitoring of a environment;
of a complete set of reagents for the bioluminescent analysis on the basis of
the isolated bienzyme system of luminous bacteria.
In the IBSO Collection, work is now underway toward
creating the database within the framework of the international program
Microbial Strain Data Network (MSDN) and the State Research - and - Development
Project "Aids to Investigations in Physico-Chemical Biology and
Biotechnology", "Microbial diversity" (the part "Culture collection
of microorganisms"). The IBSO Culture Catalogue of Luminous Bacteria is
available on disk in the MS-DOS format for personal computers. The English version of the Catalogue is
Accessible through the international network MSDN (News Letters, 1994;
Kalakoutskii et al., 1996; Rodicheva et al., 1996). The IBSO Collection designs
the database about features of natural and transgenic bacteria carring marker lux-gene (Medvedeva et al., 2001).
Participation of the Collection in the MSDN makes its activity more effective
and information exchange more expeditions.
Luminous bacteria are a convenient object to be used
in solving general biological problems, genetic and genetic-engineering
researchs. The collection of luminous bacteria can serve as a basis for
researching the strain-superproducers of luciferases, NADH:FMN-oxidoreductase,
decarboxylase, endonuclease of restriction, chitinase producers [Ruzgiene et
al, 1990, 1991, 1992, Kolykhalov et al, 1991, Repin et al., 1993, 1995a,b,
1997a,b, Vydryakova et al., 1993]. They can be used successfully to determine
toxicity of microquantity of different substances [Kratasyuk, Gitelson, 1987,
Popova et al., 1993, Kudryasheva et al., 1996].
Strains from the luminous bacteria collection are used
as a basis for preparation of lyophilized bacteria for measurements of the
integral biological toxicity of different substances: heavy metals, phenols,
narcotics, toxins, pesticides, medicines etc.
The method is based on quenching the bioluminescence of luminous
bacteria by different toxicants.
The microbiosensor based on freeze-dried luminous
bacteria has been used successfully to control the water quality in different water
bodies, such as rivers, waste water of wood-pulp and paper production
facilities and other industries [Stom et al., 1986, 1992, Popova et al., 1993,
Geel et al., 1993, Kudryasheva et al, 1996, Kuznetsov et al., 1990, 1997-2000,
Kratasyuk et al., 1991, 1997, 1998, 2001]. The preparation produced on the
basis of freeze-dried luminous bacteria in comparable with the
"Microtox" preparation produced by the Beckman but less expensive.
Unconventional techniques for cultivating of luminous
bacteria [Rodicheva et al., 1973, Zavoruev et al., 1982, Gitelson et al., 1984]
and deriving highly purified bacterial luciferases from four species (Photobacterium phosphoreum, Photobacterium
leiognathi, Vibrio fischeri and Vibrio harveyi) have been developed
[Mezhevikin et al, 1983, Bondar et al., 1988, Tyulkova, 1990, Sukovataya,
At present, biotechnology department produces sets of
analytical luminescence reagents based on the coupled enzyme system: luciferase
- NAD(P)H:FMN-oxidoreductase isolated from luminous bacteria [Tyulkova,
Illarionov, 1994, Kuznetsov et al., 1997]. A kit of reagents for analytical
bioluminescence (KRAB) can be used in medicine, ecological monitoring, and for
quality inspection of newly synthesized substances.
The diagnostic kit (KRAB) can be used for
determination of NAD-dependent dehydrogenases, proteases and antiproteases in
blood plasma and serum, platinoids etc [Petushkov et al., 1983, 1984, 1985,
1987, Kratasyuk et al., 1985, Gitelson et al., 1985, Kratasyuk, 1995].
Institute of Biophysics offers an immobilized reagent for bioluminescent assay (IRBA) [Kratasyuk et al., 1994]. Using
the kit of reagents for analytical bioluminescence (KRAB) and the hydrogenase
of hydrogen bacteria, researchers of the Institute of Biophysics have developed
methods for determining hydrogenase activity and the concentration of dissolved
hydrogen [Petushkov V.N., Guseinov O.A., 1992].
Researchers of the Institute of Biophysics have
developed the bioluminometers, a devices for measuring luminescence intensity. Such devices are now produced by the
Institute together with the "Nauka" Special Design and Development
Bacterial bioluminescence can be used as a basis for
making visual aids and creating tasks for laboratory work at schools and
universities. Bioluminescence methods may be employed in investigating fine
details of gene structure and functioning, conjugated chains of enzymatic
reactions, determining the quantity of substrates, cofactors, etc. [Andrade,
1995; Kratasyuk et al., 1995, 1997, Kratasuyk, Kudinova, 1999]. Besides its
evident educational advantages, the cold luminescence of life is aesthetically
and emotionally attractive.
Remarkable opportunities offered by the use of
luminous bacteria in the various areas of biology, their ecological and
technological applications permit us to state that they deserve close attention
of biologists and other experts of the adjacent areas of knowledge.