[Bion-M1. Biological activities of microorganisms under the conditions of a 30-day space flight].

It was stated that spaceflight factors (SFF) affect the chromosomal DNA interchange during Streptomyces crossing. Cross polarity and primary input of a parent chromosome fragment in recombinant generation imply a more lasting cells contact in microgravity and a broader horizontal transport of genetic material. SFF had no effect on recombination frequency and mutation in a model of parental auxotrophic markers reversion to prototrophism. It was demonstrated that SFF boosted the fC31 phage exit from S. lividans 66 (fC31) and did not influence phage induction in S. coelicolor A3(2) (fC31). SFF inhibited synthesis of antiobiotic actinorhodin in lisogenic S. coelicolor A3(2), and tylosin and desmicosin in S. fradiae. Survivability of electrogenic bacteria Shewanella oneidensis MR-1 in space flight was higher compared with the synchronous control experiment. The reduction activity of S. oneidensis MR-1 as an indicator of electron generation effectiveness was identical in flight and laboratory samples.

[Study of some aspects of the mechanism of bacterial synthesis of silver sulfide nanoparticles by mMetal-reducing bacteria Shewanella oneidensis MR-1].

In the present work it was shown that biosynthesis of silver sulfide nanoparticles from silver nitrate and sodium thiosulfate solutions of millimolar concentration occurs efficiently by living Shewanella oneidensis MR-1 cells, as well as by ultrasonically-disrupted cells and by the membrane fraction of the cells. The size of nanoparticles synthesized in the presence of living cells was 7.8 ± 1.5 nm, while in the presence of ultrasonically-disrupted cells--it was 6.52 nm. The shape of nanoparticles in both cases was close to spherical. It was also shown, that synthesis of nanoparticles occurs in a cell-free solution of sodium thiosulfate that has been incubated with cells previously and to which then a silver nitrate solution was added. In this case the nanoparticles were of elongated shape and their size was (11 ± 4) x (24 ± 6) nm. In the control experiment, when only silver nitrate and sodium thiosulfate solutions not incubated with cells were used, the nanoparticles were not detected. It was shown that biosynthesis of nanoparticles occurs both in aerobic and anaerobic conditions. Nanoparticles are not formed by using thermally inactivated cells as it was shown by us previously. The results show the important role of the native structures of cells for the nanoparticles formation.

[Serological detection of Rhesus antigen D in blood traces from Rh-negative (cde) people]. / O serologicheski vyiavliaemom nalichii v sledakh krovi Rezus-otritsatel'nykh (cde) lits Rhesus-antigena D.

A D-like structure was serologically detected on the internal side of Rh-negative (cde) erythrocyte membrane. This structure is absolutely inaccessible for anti-D antibodies in morphologically intact liquid blood erythrocytes. In hemolyzed erythrocytes of blood stains this antigenic structure, serologically identified as Rh antigen D, is accessible for binding with anti-D antibodies and for detection by the absorption-elution test, particularly so after blood stain treatment with proteases.

[Biosynthesis of leucine aminopeptidase by Xanthomonas rubrilineans 67 in culture with various concentrations of nitrogen containing compounds]. / Biosintez leitsinaminopeptidazy kul'turoi Xanthomonas rubrilineans 67 pri var'irovanii azotosoderzhashchikh komponentov sredy.

The dynamics of consumption of amine and ammonium nitrogen and glucose in the process of Xanthomonas rubrilineans 67 growth and biosynthesis of leucine aminopeptidase was studied. It was shown that the rate of leucine, alanine and glycine consumption as a source of amine nitrogen out of 16 amino acids was the highest during the fermentation. Addition of these three amino acids or their mixtures to the medium at definite stages of the fermentation process increased the leucine aminopeptidase biosynthesis by 50 to 100 per cent. Ammonium nitrogen was not used by X.rubrilineans 67. The consumption of glucose during the fermentation was even: by the 24th hour of the process the medium contained about 10 per cent of the glucose initial concentration. The optimal temperature for the culture growth and leucine aminopeptidase biosynthesis was determined. It was shown to be 28 degrees C. Higher aeration increased the culture productivity.

[Isolation of protoplasts of Xanthomonas rubrilineans and their use in the study of localization of aminopeptidases]. / Poluchenie protoplastov Xanthomonas rubrilineans i ikh ispol'zovanie dlia izucheniia lokalizatsii aminopeptidaz.

The culture of Xanthomonas rubrilineans was able to synthesize a number of intracellular aminopeptidases. To study localization of the enzymes in the cells, a protoplasting procedure was developed providing the yield of 99.7 per cent. The following subcellular fractions were isolated: periplasmic, cytoplasmic and membranous. It was shown that alanine aminopeptidase was a cytoplasmic enzyme and glutamate peptidase was a membrane-bound enzyme.

[Relation between the morphogenesis of Xanthomonas rubrilineans and biosynthesis of aminopeptidase in the process of growth and development of its producer]. / Vzaimosviaz' morfogeneza kul'tury Xanthomonas rubrilineans i biosinteza aminopeptidazy v protsesse rosta i razvitiia produtsenta.

The dynamics of growth and development of Xanthomonas rubrilineans, a culture producing intracellular aminopeptidase, was studied. A difference between the growth rate determined by intensity of the total biomass accumulation and the rate of the culture multiplication was found. The difference was due to the presence of two phases in the culture development during the exponential growth: the phase of increasing the linear sizes of the cells and the phase of the culture intensive multiplication. The most intensive synthesis of aminopeptidase was observed during the phase of increasing the linear sizes of the cells. The dynamics of consumption of the main sources of carbon and nitrogen by the culture was investigated.

[Experimental study of protease C--proteolytic enzyme of microbial origin]. / Eksperimental'noe izuchenie proteazy C--proteoliticheskogo fermenta mikrobnogo proiskhozhdeniia.

The specific activity of protease C, a proteolytic enzyme isolated from Acremonium chrysogenum was studied under experimental conditions. Protease C was shown to lyse necrotic biological substrates (dry crusts of burn wounds) and blood clots. By the nature of the effect protease C was analogous to terrilytin and by the level of the effect it was superior in some experiments. Protease C was low toxic and had no mutagenic action.

[Intracellular aminopeptidase from Xanthomonas rubrilineans, hydrolyzing alpha-amino acid esters and cefalexin]. / Vnutrikletochnaia aminopeptidaza iz Xanthomonas rubrilineans, gidrolizuiushchaia éfiry alpha-aminokislot i tsefaleksin.

The aminopeptidase was isolated from cell-free extracts of Xanthomonas rubrilineans by protein precipitation by isopropyl ester with subsequent purification by affinity chromatography on CABS-Sepharose, bacitracin-Sepharose, gel filtration through Sephadex G-200 and ultrafiltration, the total yield being 32% with 2200-fold purification. The enzyme was homogeneous during SDS-PAAG electrophoresis. Apart from the broad spectrum of the peptidase activity, aminopeptidase possesses a hydrolase activity towards beta-lactam antibiotics and an esterase activity towards L- and D-amino acids. Besides, this enzyme catalyzes the acetyl transfer reaction during cephalexin synthesis from the D-phenylglycine ester and 7-aminodesacetoxycephalosporanic acid. The maximal enzyme activity during L-Ala-pNA and cephalexin hydrolysis is manifested at pH 6.5. The enzyme is stable at pH 4.0-8.0 and is inhibited by o-phenanthroline, p-chloromercuribenzoate, hydrogen acetate and N-bromosuccinimide. The molecular mass of the enzyme is 270-280 kDa. The enzyme is a tetramer; the molecular mass of each of its four subunits is 70 +/- 2 kDa. The isoelectric point for the enzyme is 6.8. The amino acid composition of the enzyme appears as follows: Asp63, Thr33, Ser32, Glu72, Gly55, 1/2Cys3-4, Val45, Ile24, Leu53, Tyr23, Phe24, Lys23, His16, Arg36, Pro60, Met25, Ala55.

[Differentiation of the fungus Acremonium chrysogenum and the synthesis of serine proteinases]. / Differentsiatsiia griba Acremonium chrysogenum i sintez serinovykh proteinaz.

The biosynthesis of serine proteinases by the fungus Acremonium chrysogenum was studied in the process of its growth in media differing in the content of a protein substrate. Morphological differentiation of the submerged fungal culture was shown to be characterised by two reproduction pathways (conidiogenesis and arthrosporogenesis) and by the corresponding synthesis of serine proteinases II and I. The synthesis of serine proteinase I and cephalosporin was found to correlate with the polycyclic culture growth caused by the formation and germination of single spherical arthrospores.

[Enzyme preparations for research on protein hydrolysates and amino acid mixtures not containing peptides]. / Fermentnye preparaty dlia izucheniia belkovykh gidrolizatov i smesei aminokilsot, ne soderzhashchikh peptidov.

Some properties and the hydrolysing ability of two novel enzyme preparations, a proteolytic preparation "C" from Acremonium chrysogenum (Cephalosporium acremonium) and a peptidase preparation (the producer from the family Pseudomanadaceae), are described. The preparations can be used for obtaining protein hydrolysates with different ratios of free amino acids and peptides. The protein hydrolysis with the preparation "C" enables one to obtain hydrolysates containing 13-18% of free amino acids. The further treatment of the hydrolysates with the peptidase preparation results either in complete hydrolysis of the remained peptide fractions or in obtainment of solutions containing from 60 to 85% of free amino acids and low-molecular weight peptides.

[Isoelectric focusing of a preparation of proteolytic enzymes from Streptomyces 771]. / Izoélektricheskoe fokusirovanie preparata proteoliticheskikh fermentov iz Streptomyces 771.

The proteolytic enzymes contained in the preparation from Streptomyces 771 have been separated by isoelectric focusing in the sucrose density gradient at pH 3-10. The following enzymes have been identified: three multiple forms of neutral metal proteinase (pI 5.1, 6.37, 7.8) each of which splits DNP-Gly-Gly decreases-Val-ArgOMe; elastase-like metal proteinase active with respect to RBB-elestin with pI 10.68; metal-dependent peptidases: leucin aminopeptidase active with respect to L-RBB-elastin with pI 10.68 metal-dependent peptidases: leucin aminopeptidase active with respect to L-leucin n-nitroanilide and L-leucin beta-naphtylamide with pI 7.65, 7.15, 6.67, 6.45, 5.7, 5.35, 5.22, 4.83; carboxy peptidase with pI 5.95, 6.37; serine metal-dependent subtilisin-like proteinase active with respect to 2-Ala-Ala-LeupNA, 2-Gly-Gly-LeupNA, 2-Ala-LeupNA; two multiple forms of serine trypsin-like proteinase active with respect to BAEE and BApNA with pI 4.35, 4.76; serine chymotrypsin-like proteinase with pI 8.68 active with respect to ATEE.