The text of the original in Russian 


DEVELOPMENT OF RAPID ASSESSMENT METHOD FOR INCLUSION OF DEUTERIUM AND CARBON D C13 in the molecule of amino acids OF BACTERIAL 


We propose to use highly mass spectrometry electron impact of volatile derivatives of amino acids for the rapid assessment of the level of incorporation of stable isotopes of deuterium and carbon 13 C D in molecules secreted amino acids and amino acid residues of proteins of total biomass of L-strain fenilalaninprodutsiruyuschego optional metilotrofnyh bacteria Brevibacterium methylicum, the assimilation of methanol ribulozo - 5-way monofosfatnomu carbon fixation, and L-leytsinprodutsiruyuschego strain of bacteria obligate metilotrofnyh Methylobacillus flagellatum, realizing 2-keto-3-dezoksiglyukonataldolazny option ribulozo-5-monofosfatnogo ways of carbon fixation when growing the bacteria in medium containing a source of stable isotopes (D) methanol (13 C) methanol and heavy water. Also investigated the inclusion of L-[2,3,4,5,6-D] phenylalanine, L-[3,5-D] tyrosine and L-[2,4,5,6,7-D] tryptophan in the membrane protein bakteriorodopsin SYNTHESIZED Halobacterium halobium ET 1001. For mass spectrometric analysis of multicomponent mixtures of amino acids in the composition of lyophilized culture fluids and protein hydrolysates were subjected sparing hydrolysis in 6 M DSl (3% phenol) and 2 M barium hydroxide, followed by modification of amino acids and methyl esters of N-dimetilaminonaftalin-5-sulfonil-derived amino acids that preparative razdelelyali treatment-phase high performance liquid chromatography and then analyzed by mass spectrometry electron impact EI on the device "MB-80 A" (Hitachi, Japan) with the energy of ionizing electrons 70 eV. The resulting molecule isotope-labeled [D] - and [13C] amino acids were a mixture of varying the number of atoms in a molecule isotopes. Levels of incorporation of 13 C and D molecules secreted amino acids and amino acid residues of proteins of total biomass is investigated using this method vary depending on the content of tracer substrates into the growth media, and different for different amino acids and 10% for L-leytsina/izoleytsina and to 97.5% for L-alanine. Therefore, as the proposed method can be used for mass spectrometric analysis of rapid incorporation of stable isotopes of atoms in the molecule synthesized amino acids. 

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

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