European Journal of Molecular Biotechnology

58-76

Authors: Oleg Mosin, Ignat Ignatov

By the method of electron impact mass-spectrometry was studied the pathways of biosynthesis of 2H, 13C-labeled amino acids of a facultative methylotrophic bacterium Brevibacterium methylicum and an obligate methylotrophic bacterium Methylobacillus flagellatum obtained on growth media containing as a source of stable isotopes [2H]methanol, [13C]methanol and 2H2O. For mass-spectrometric analysis the multicomponential mixtures of 2H- and 13C-labeled amino acids, derived from cultural media and protein hydrolysates after hydrolysis in 6 M 2HСl (3 % phenol) and 2 M Ва(OH)2 were modified into N-benzyloxycarbonyl-derivatives of amino acids as well as into methyl esters of N-5-(dimethylamino)naphthalene-1-sulfonyl chloride (dansyl) derivatives of [2H, 13С]amino acids, which were preparative separated using a method of reverse-phase HCLP. Biosynthetically obtained 2H- and 13C-labeled amino acids represented the mixtures differing in quantities of isotopes incorporated into molecule. The levels of 2H and 13С enrichment of secreted amino acids and amino acid resigues of protein were found to vary from 20,0 atom % to L-leucine/isoleucine up to 97,5 atom % for L-alanine depending on concentration of 2H- and 13C-labelled substrates.

85-90

Authors: Natalya A. Sidorova, Philip D. Voronov

The article presents the results of a study of mutagenic effects of Pseudomonas alcaligenеs polysaccharides. Pseudomonas genus – non-fermentative ubiquitous bacteria, having specific metabolic cycles and unique physical, chemical and biological properties was used as a producer of natural exopolysaccharides. In an experiment using the Ames test, three variants of test compounds were studied: 1. a compound of the Pseudomonas alcaligenes biofilm, 2. exopolysaccharide matrix and the microorganism cell wall compound, and 3. actually the microbial exopolysaccharide. In all cases the lack of mutagen action of polysaccharides of Pseudomonas alcaligenes is proved that make them perspective for use as nanomaterials of new generation – alternative wound coverings.