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Journal №6 for 2014 г.
Article in number:
Biosynthesis of deuterium labeled purine ribonucleoside riboxine with high levels of isotopic enrichment by chemo heterotrophic bacterium Bacillus subtilis
Keywords: Bacillus subtilis [<sup>2</supH]riboxine biosynthesis heavy water fast atom bombardment mass spectrometry (FAB)
Authors:
О.V. Mosin - Ph.D. (Chem.), Research Scientict, Moscow State University of Applied Biotechnology, Russia. E-mail: mosin-oleg@yandex.ru
I.I. Ignatov - D.Sc., Doctor of European Academy of Natural Sciences (Hanover, Germany), Professor, Director of Scientific-Research Center of Medical Biophysics (София), Bulgaria. E-mail: mbioph@dir.bg
Abstract:
Microbial synthesis of 2H-labeled purine ribonucleoside riboxine (yeald 3,9 g/l) with using an adapted to deuterium strain of Gram-positive chemo heterotrophic bacteria Bacillus subtilis on heavy water medium with highly level of deuteration (99,8 at.% 2H2O) with 2% hydrolysate of deuterated biomass of methylotrophic bacterium Brevibacterium methylicum VKPM В-3157 as a source of 2H-labeled growth substrates, obtained from the minimal М9 growth medium with 98 % 2Н2О and 2 % [2H]methanol was carried out. Fractionation of riboxine was carried out by adsorption/resorption on a surface of activated carbon coal, extraction by 0,3 M NH4-formiate buffer (рН = 8,9) with the subsequent cristallization in 80% of ethanol and column ion exchange chromatography on cation exchanger AG50WX 4, counterbalanced with 0,3 M NH4-formiate with 0,045 M NH4Cl. The level of deuterium enrichment of [2H]riboxine, analyzed by a method of FAB mass-spectrometry makes up 5 deuterium atoms with incorporation of 3 deuterium atoms into ribose and 2 deuterium atoms into hypoxantine fragments of the molecule.
Pages: 34-45
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