Abstract
We used Gram-positive aerobic facultative methylotrophic bacterium, Brevibacterium methylicum, L-phenylalanine producer with ribulose-5-monophosphate (RuMP) cycle for carbon assimilation for microbiological preparation of [ 2 H]phenylalanine via conversion of low molecular weight substrates ([U- 2 H]MeOH and 2 H2O). For this purpose, the cells of the methylotroph with improved growth characteristics were used on minimal salt media M9 supplemented with 2 % (v/v) [U- 2 H]MeOH and increasing gradient of 2 Н2O concentration from 0; 24,5; 49,0; 73,5 up to 98 % (v/v) 2 Н2O. L-phenylalanine was isolated from the growth medium after adding 5 M 2 HCl (in 2 Н2О), pH = 2,0 by extraction with isopropanol and subsequent crystallization in ethanol (output 0,65 g/l). Alanine, valine, and leucine/isoleucine were produced and accumulated exogenously in amounts of 5–6 mol in addition to the main product of biosynthesis. The method allows to obtain [ 2 Н]amino acids with different levels of deuterium enrichment, depending on 2 Н2O concentration
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