title: [Microbiological synthesis of [2H]-inosine with a high degree of isotopic enrichment by the gram-positive chemoheterotrophic bacterium Bacillus subtilis].
abstract: A 2H-labeled purine ribonucleoside inosine was microbiologically synthesized (yield, 3.9 g/L of culture liquid) using a deuterium-adapted strain ofthe gram-positive chemoheterotrophic bacterium Bacillus subtilis, cultivated in a heavy water medium with a high degree of deuteration (99.8 at % 2H) containing 2% hydrolysate of deuterated biomass of the methylotrophic bacterium Brevibacterium methylicum as a source of 2H-labeled growth substrate produced in an M9 minimal medium with 98% 2H20 and 2% [2H]-methanol. The inosine extracted from the culture liquid of the producer strain was fractionated by adsorption (desorption) on an activated carbon surface, extraction with 0.3 M ammonium-formate buffer (pH 8.9), subsequent crystallization in 80% ethanol, and ion exchange chromatography on a column with AG50WX 4 cation exchange resin equilibrated with 0.3 M ammonium-formate buffer containing 0.045 M NH4Cl. Fast atom bombardment (FAB) mass spectrometry demonstrated incorporation of five deuterium atoms in the inosine molecule (62.5% 2H), three of which were contained in the ribose moiety and two in the hypoxanthine moiety.
authors.forename: O V
authors.initials: OV
authors.lastname: Mosin
authors.forename: V I
authors.initials: VI
authors.lastname: Shvets
authors.forename: D A
authors.initials: DA
authors.lastname: Skladnev
authors.forename: I
authors.initials: I
authors.lastname: Ignatov
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry*; D001951:Brevibacterium / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry* / Q000378:metabolism; D006094:Gram-Positive Bacteria; D007288:Inosine / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D016339:Spectrometry, Mass, Fast Atom Bombardment
chemical_list: D007288:Inosine; D003903:Deuterium