Microbiological Synthesis of 2 Н- and 13 С-Labeled Amino Acids and Proteins With Various Levels of Isotopic Enrichment

By the method of microbiological synthesis were obtained and analyzed by electron impact mass-spectrometry the evaluation of 2 H and 13 C enrichment levels of secreted amino acids of methylotrophic bacteria Brevibacterium methylicum and Methylobacillus flagellatum, and amino acid resigues of total protein obtained from media contaning as a sourse of stable isotopes [ 2 H]methanol, [ 13 C]methanol and 2 H2O. It was also performed the incorporation of L-[2,3,4,5,62 Н]phenylalanine, L-[3,5- 2 Н]tyrosine and L-[2,4,5,6,7- 2 Н]tryptopan in bacteriorhodopsin synthesised in purple membranes of photo-organotrophic halobacterium Halobacterium halobium ET 1001. For mass-spectrometric analysis the multicomponential mixures of 2 H- and 13 C-labeled amino acids, derived from cultural media and protein hydrolysates after hydrolysis in 6 M 2 HСl (3 % phenol) and 2 M Ва(OH)2 were modified into N-benzyloxycarbonyl-derivatives of amino acids as well in methyl esters of N-dansyl-derivatives of amino acids, which were preparative separated using a method of reverse-phase HCLP. 2 H- and 13 C-labeled amino acids represented the mixures differing in quantities of isotopes incorporated into molecule. The levels of 2 H and 13 С enrichment of secreted amino acids and amino acid resigues of protein were found to vary from 20,0 atom % to