The biological effects of heavy water prove to be very interesting question of nowdays biochemical science which comprising two main points: the evolution of deuterium itself as well as the chemical processes going on with participation of deuterium oxyde.
The Kirlian effect in which a Kirlian aura is observed is called the plasma emission of light of the electric discharge. Kirlian effect is color coronal discharge in gas. The discharge is on the surface of the objects located in an alternating electric field with high frequency from 10 to 100 kilohertz. There occurs a surface interaction of 5 to 30 kW between the electrode and the object under research.
The biosynthesis of 2H-labeled phenylalanine was done by converse of low molecular weight substrates ([U2H]methanol and 2H2O) in a new RuMP facultative methylotrophic mutant Brevibacterium methylicum. To make the process work, adapted cells with improved growth characteristics were used on minimal medium M9 with the maximum content of 2H-labeled substrates. Alanine, valine, and leucine/isoleucine were produced and accumulated exogeneously in addition to the main product of biosynthesis. Electron impact mass spectrometry of methyl esters of the N-Dns-amino acid mixture obtained after the chemical derivatization of growth medium with dansyl chloride and diazomethane, was done to calculate the deuterium enrichment of the amino acids synthesized. The experimental data testified to the character of labeling of amino acid molecules as heterogeneous; however, high levels of deuterium enrichment were detected in all presented molecules - for phenylalanine the enrichment was six, leucine/isoleucine - 5.1, valine - 4.7, and alanine - 3.1 deuterium atoms.
It was proposed to use the 2H-labeled hydrolysate of RuMP facultative methylotroph Brevibacterium methylicum, obtained from deuterated salt medium dM9 as a substrate for the growth of inosine producing bacterium Bacillus subtilis. The growth of the bacterim was performed via glucose convertion on specially developed medium dHM with 78.5% (m/m) 2H2O and supplimented with 2.5% (m/m) of 2H-labeled methylotrophic hydrolysate. To evaluate the level of deuterium enrichment FAB MS technique was used after the isolation of 2H-labeled inosine. 2H-labeled inosine obtained from dHM medium represented a mixture of molecular species containing various number of included deuterium atoms with different contribution to the enrichment. The level of enrichmet calculated by the presence of most abandant peak of the molecular ion in cluster ((M+H)+ at m/z 274) was estimated as five deuterium atoms, from which three are attributed to ribose and two to hypoxantine.
The physiological adaptation process of various bacterial producents of amino acids, proteins and nucleosides belonging to various taxsonomic groups of microorganisms (facultative and obligate methylotrophic bacteria - Brevibacterium methylicum and Methylobacills flagellatum, halophilic bacterium - Halobacterium halobium and bacills - Bacillus subtilis) to growth and biosynthesis of necessary compounds on media containing the maximum concentration of heavy water is investigated. In article is informed on a method, which consists in multistep adaptation of bacteria to deuterium with the folowing selection of individual colonies grown on heavy water. In the result of application of the given approach among the studied bacteria were selected the individual strains keeping high growth and biosynthetic abilities while growing on heavy water.
Department of Biotechnology, M. V. Lomonosov State Academy of Fine Chemical Technology, Vernadskogo Prospekt 86, 117571, Moscow, Russia
ABSTRACT The biological effects of heavy water prove to be very interesting question of nowdays biochemical science which comprising two main points: the evolution of deuterium itself as well as the chemical processes going on with participation of deuterium oxyde.