Experimental data on size distribution of micro- and macroaggregates and primary soil particles were obtained by laser diffraction analysis of the samples of humus horizons of the zonal soil...

The oldest prokaryotic photoautotrophic organisms, cyanobacteria, produce many different metabolites. Among them is the water-soluble neurotoxic non-protein amino acid beta-N-methylamino-L-alanine (BMAA), whose biological functions in cyanobacterial metabolism are of fundamental scientific and practical interest. An early BMAA inhibitory effect on nitrogen fixation and heterocyst differentiation was shown in strains of diazotrophic cyanobacteria Nostoc sp. PCC 7120, Nostoc punctiforme PCC 73102 (ATCC 29133), and Nostoc sp. strain 8963 under conditions of nitrogen starvation. Herein, we present a comprehensive proteomic study of Nostoc (also called Anabaena) sp. PCC 7120 in the heterocyst formation stage affecting by BMAA treatment under nitrogen starvation conditions. BMAA disturbs proteins involved in nitrogen and carbon metabolic pathways, which are tightly co-regulated in cyanobacteria cells. The presented evidence shows that exogenous BMAA affects a key nitrogen regulatory protein, PII (GlnB), and some of its protein partners, as well as glutamyl-tRNA synthetase gltX and other proteins that are involved in protein synthesis, heterocyst differentiation, and nitrogen metabolism. By taking into account the important regulatory role of PII, it becomes clear that BMAA has a severe negative impact on the carbon and nitrogen metabolism of starving Nostoc sp. PCC 7120 cells. BMAA disturbs carbon fixation and the carbon dioxide concentrating mechanism, photosynthesis, and amino acid metabolism. Stress response proteins and DNA repair enzymes are upregulated in the presence of BMAA, clearly indicating severe intracellular stress. This is the first proteomic study of the effects of BMAA on diazotrophic starving cyanobacteria cells, allowing a deeper insight into the regulation of the intracellular metabolism of cyanobacteria by this non-protein amino acid.

All cyanobacteria produce a neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA). However, the biological function of BMAA in the regulation of cyanobacteria metabolism still remains undetermined. It is known that BMAA suppresses the formation of heterocysts in diazotrophic cyanobacteria under nitrogen starvation conditions, and BMAA induces the formation of heterocyst-like cells under nitrogen excess conditions, by causing the expression of heterocyst-specific genes that are usually “silent” under nitrogen-replete conditions, as if these bacteria receive a nitrogen deficiency intracellular molecular signal. In order to find out the molecular mechanisms underlying this unexpected BMAA effect, we studied the proteome of cyanobacterium Nostoc sp. PCC 7120 grown under BMAA treatment in nitrogen-replete medium. Experiments were performed in two experimental settings: (1) in control samples consisted of cells grown without the BMAA treatment and (2) the treated samples consisted of cells grown with addition of an aqueous solution of BMAA (20 µM). In total, 1567 different proteins of Nostoc sp. PCC 7120 were identified by LC-MS/MS spectrometry. Among them, 80 proteins belonging to different functional categories were chosen for further functional analysis and interpretation of obtained proteomic data. Here, we provide the evidence that a pleiotropic regulatory effect of BMAA on the proteome of cyanobacterium was largely different under conditions of nitrogen-excess compared to its effect under nitrogen starvation conditions (that was studied in our previous work). The most significant difference in proteome expression between the BMAA-treated and untreated samples under different growth conditions was detected in key regulatory protein PII (GlnB). BMAA downregulates protein PII in nitrogen-starved cells and upregulates this protein in nitrogen-replete conditions. PII protein is a key signal transduction protein and the change in its regulation leads to the change of many other regulatory proteins, including different transcriptional factors, enzymes and transporters. Complex changes in key metabolic and regulatory proteins (RbcL, RbcS, Rca, CmpA, GltS, NodM, thioredoxin 1, RpbD, ClpP, MinD, RecA, etc.), detected in this experimental study, could be a reason for the appearance of the “starvation” state in nitrogen-replete conditions in the presence of BMAA. In addition, 15 proteins identified in this study are encoded by genes, which are under the control of NtcA—a global transcriptional regulator—one of the main protein partners and transcriptional regulators of PII protein. Thereby, this proteomic study gives a possible explanation of cyanobacterium starvation under nitrogen-replete conditions and BMAA treatment. It allows to take a closer look at the regulation of cyanobacteria metabolism affected by this cyanotoxin.

Due to the emergence of multiple antibiotic resistance in many pathogens, the studies on new antimicrobial peptides (AMPs) have become a priority scientific direction in fundamental and applied biology. Diverse mechanisms underlie the antibacterial action of AMPs. Among them are the effects that AMPs cause on bacterial cell membranes. In this work, we studied the antibacterial activity of a peptide named P4 with the following sequence RTKLWEMLVELGNMDKAVKLWRKLKR that was constructed from two alpha-helical fragments of the influenza virus protein M1 and containing two cholesterol-recognizing amino-acid consensus (CRAC) motifs. Previously we have shown that 50 μM of peptide P4 is toxic to cultured mouse macrophages. In the present work, we have found that peptide P4 inhibits the growth of E. coli and B. subtilis strains at concentrations that are significantly lower than the cytotoxic concentration that was found for macrophages. The half-maximal inhibitory concentration (IC50) for B. subtilis and E. coli cells were 0.07 ± 0.01 μM and 1.9 ± 0.4 μM, respectively. Scramble peptide without CRAC motifs did not inhibit the growth of E. coli cells and was not cytotoxic for macrophages but had an inhibitory effect on the growth of B. subtilis cells (IC50 0.4 ± 0.2 μM). A possible involvement of CRAC motifs and membrane sterols in the mechanism of the antimicrobial action of the P4 peptide is discussed. We assume that in the case of the Gram-negative bacterium E. coli, the mechanism of the toxic action of peptide P4 is related to the interaction of CRAC motifs with sterols that are present in the bacterial membrane, whereas in the case of the Gram-positive bacterium B. subtilis, which lacks sterols, the toxic action of peptide P4 is based on membrane permeabilization through the interaction of the peptide cationic domain and anionic lipids of the bacterial membrane. Whatever the mechanism can be, we report antimicrobial activity of the peptide P4 against the representatives of Gram-positive (B. subtilis) and Gram-negative (E. coli) bacteria.

The consumption specters of organic matter fractions in ultraviolet and visible parts of specter (UV-visible specters) were investigated. The specters of soluted organic matter, as well as labile humic matters and humic matters itselves, which were extracted from small, medium and large aggregates from typical chernozems of long-term experiments were investigated. The samples of undisturbed chernozems, which are annually collected in the annually hayed steppe and from the chernozem with extreme organic matter degradation, were used (variant of long-termed experiment - irreplaceable black fallow since 1964). On the basis of specters obtained the parameters of optical indices SUVA254, E2/E3, E4/E6 were calculated. The highest differences between the aggregates were revealed in extracts of organic matter. The differences were lower for labile humic matters. There were no differences revealed for humic matters between the optical indices for different aggregates of one type of chernozems use. It is shown that all of the three parameters are not changed symbasically. Hence, all of the three parameters should be used for complete spectral characteristic.

The effect of detonation-synthesis nanodiamonds (NDs) on the viability of bacterium Rhodococcussp. 2012B and its capability for ND biodegradation was determined. It was shown that NDs did not...