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Applied Biochemistry and Microbiology

, Volume 49, Issue 3, pp 233–243 | Cite as

Microbiological synthesis of [2H]-inosine with a high degree of isotopic enrichment by the gram-positive chemoheterotrophic bacterium Bacillus subtilis

  • O. V. MosinEmail author
  • V. I. Shvets
  • D. A. Skladnev
  • I. Ignatov
Article
  • 69 Downloads

Abstract

A 2H-labeled purine ribonucleoside inosine was microbiologically synthesized (yield, 3.9 g/L of culture liquid) using a deuterium-adapted strain of the 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% 2H2O 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.

Keywords

Deuterium Apply Biochemistry Inosine Deuterium Atom Fast Atom Bombardment Mass Spectrometry 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Pleiades Publishing, Ltd. 2013

Authors and Affiliations

  • O. V. Mosin
    • 1
    Email author
  • V. I. Shvets
    • 1
  • D. A. Skladnev
    • 2
  • I. Ignatov
    • 3
  1. 1.Lomonosov State University of Fine Chemical TechnologyMoscowRussia
  2. 2.State Research Center of Genetics and Selection of Industrial MicroorganismsMoscowRussia
  3. 3.Scientific Research Center of Medical BiophysicsSofiaBulgaria

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