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Using of facultative methylotrophic bacterium Brevibacterium methylicum В-5652 with RMP-cycle of carbon assimilation for microbiological synthesis of [2H]phenylalanine with different levels of deuterium enrichment
Authors: Oleg Mosin, Ignat Ignatov, Dmitry Skladnev, Vitaly Shvets
Number of views: 371
With using of a strain of of L-phenylalanine secreted Gram-positive aerobic facultative methylotrophic bacteria Brevibacterium methylicum В-5652, assimilating methanol via ribulose-5-monophosphate (RMP) cycle of carbon assimilation it was carried out the preparative microbiological synthesis of phenylalanine and metabolically related amino acids (alanine, valine, leucine/isoleucine in the amount of 5–6 mmol/l), labelled with deuterium (2H). The data on adaptation of L-phenylalanine secreted methylotrophic bacterium В. methylicum to the maximal concentration of deuterium in minimal growth medium M9 with 98 % 2Н2O and 2 % [2Н]methanol, and data on biosynthesis of deuterium labelled L-phenylalanine with different levels of enrichment are submitted. The developed method for biosynthesis allows to obtain [2Н]phenylalanine with different levels of isotopic enrichment, depending on 2Н2O concentration in growth media M9, from 17 % (2 deuterium atoms) (on growth medium with 24,5 % 2Н2О) right up to 75 % (6 deuterium atoms) (on growth medium with 98 % 2Н2О) with introduction of deuterium to benzyl С6Н5СН2-fragment of molecule that is confirmed with the data of the electron impact (EI) mass-spectrometry analysis of methyl ethers of N-dimethylamino(naphthalene)-5-sulfochloride (dansyl) derivatives of [2Н]phenylalanine, isolated from the liquid culture after its separation by RP HPLC.