"Methylotrophic biomass as 2H-labeled substrate for biosynthesis of inosine"

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Methylotrophic biomass as 2H-labeled substrate for biosynthesis of inosine Oleg V. Mosin1 1 M. V. Lomonosov State Academy of Fine Chemical Technology, Vernadskogo Prospect 86, Moscow, 117571 Abstract 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. Keywords: 2H-labeled growth substrates -

Bacillus subtilis - Biosynthesis - 2H-labeled inosine Introduction Nucleosides labeled with deuterium (2H) and other stable isotopes are becoming an indispensable tool for biomedical diagnostic and the investigation of various aspects of the metabolism [1, 2]. Thus inosine which is known as an important intermediate in the synthesis of inosine monophosphate (IMP) is in the focal point of clinical interest in medical diagnostic of heart deceases and in certain

medical cases [3, 4]. There are several approaches reported for the preparation of 2H- nucleosides. Chemical synthesis are usually tedious and inefficient. Only by employing mutant forms of bacteria, which can produce a large quantities of the nucleosides when growing of an organism on media containing deuterated substrates, the desired biochemicals can be obtained both with high yields and enrichments.

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