The text of the original in Russian 


METHOD OF OBTAINING deuterated purine inosine RIBONUKLEOZIDA HIGH-LEVEL DEYTERIROVANNOSTI 


A method for obtaining deuterated purine inosine ribonukleozida (yield 3.9 g c 1 l growth medium), high-level deyterirovannosti (61% of hydrogen atoms in the molecule substituted for deuterium). The method was adapted to the cultivation of deuterium strain of Bacillus subtilis - producer of inosine tyazhelovodorodnoy medium with 2% by the biomass hydrolysates metilotrofnoy bacteria Brevibacterium methylicum as a source of deuterium tracer-growth substrates (conditions for obtaining hydrolysates: autoclaving deuterium-biomass in a 0.1. DSl 30 -- 40 min at 08 AIT). Conditions for obtaining deuterated inosine: tyazhelovodorodnaya synthetic medium with 99.9%-term heavy water (mass%): glucose  12; D-tracer hydrolysates B. methylicum  2,  2, ammonium nitrate, magnesium sulfate  1; calcium carbonate - 2. Cultivation of bacteria was carried out in flasks Erlenmeyera with a capacity of 500 ml (filled to 100 ml) for 5-6 days at 30-320p in the conditions of intensive aeration. Allocation of deuterated inosine was in the low-temperature deposition of high impurity organic solvents acetone and methanol, solid-phase adsorption / desorption on the surface of activated carbon, extraction extractive product, and recrystallization of ion-exchange chromatography. Proteins and polysaccharides were removed nizkotemperatrnym deposition of acetone at-40C, in the subsequent adsorption of total ribonukleozidov activated carbon in the cold. Desorbirovannye ribonukleozidy derived from reacted solid phase elyutsiey ethanol-ammonia solution with the 600S, and the extraction of inosine 0.3 M ammonium formiatnym buffer (pH 8.9), followed by recrystallization in 80% ethanol. The final stage of purification was the ion exchange column chromatography on kationoobmennike AG50WX 4, balanced 0.3 M ammonium formiatnym buffer with 0.045 M ammonium chloride in a izokraticheskoy elyutsii with TLC control at R = 0.5 (yield 3.9 g c 1 l growth medium). Isotopic composition obtained inosine, investigated by means of mass spectrometry in pulsed FAB mass spectrometer VG-70 SEQ (Fisons, VG Analitical, USA) equipped with tsezievym source of Cs + on glitserinovoy matrix of the acceleration voltage of 5 kV and an ion current of 0.6-0.8 mA, showed peaks of including 4 at. deuterium (20%), 5 at. deuterium (38%), 6 atoms of deuterium (28%), and 7 atoms of deuterium (14%) with the inclusion of deuterium in ribozny and fragments of molecules gipoksantinovy inosine. 

Moscow State Academy of Fine Chemical Technology. MV Lomonosov Moscow State University, 117571 Moscow, prosp. Vernadskogo, 86. 


References 

1. Shvets VI, Yurkevich AM, Mosin OV, Skladnev DA / / Karadeniz Journal of Medical Sciences, 1995. V. 8. N 4. P. 231. 
2. Mosin OV, folding DA, Egorova TA, Shvets VI / / Biotechnology. 1996. Number 10. S. 24-40. 
3. Mosin OV, folding, DA, Shvets VI / / Izv. Russian Academy of Sciences. Ser. Biol. N 3. Pp 1-10