The dataset on the draft whole-genome sequences of two Pseudomonas aeruginosa strains isolated from urine samples of patients with urinary tract diseases.

Pseudomonas aeruginosa is a widespread multidrug-resistant opportunistic human pathogen with an extremely high mortality rate that leads to urinary tract infection morbidities in particular. Variability and dynamics in genome features and ecological flexibility help these bacteria adapt to many environments and hosts and underlie their broad antibiotic resistance. Overall, studies aimed at obtaining a deeper understanding of the genome organization of UTI-associated P. aeruginosa strains are of high importance for sustainable health care worldwide. Herein, we report the draft assembly of entire genomes of two P. aeruginosa strains, PA18 and PA23, isolated from voided urine of patients with urinary tract diseases (hydronephrosis and urolithiasis, respectively) and determine the most important genetic features for pathogenesis and virulence. Whole-genome sequencing and annotation of genomes revealed high similarity between the two UTI strains along with differences in comparison with other uropathogenic P. aeruginosa and reference strains. The 6 981 635 bp and 6 948 153 bp draft genome sequences with GC contents of 65.9% and 65.8%, respectively, provide new insights into the virulence genetic factors and genes associated with antimicrobial resistance. The whole genome data of PA18 and PA23 have been deposited in the NCBI GenBank database (accession numbers JAQRBF000000000.1 and JAQRBG000000000.1, respectively).

Features of Hemolysin Biosynthesis by Morganella morganii.

We studied the influence of medium composition and aeration on the hemolytic activity of uropathogenic Morganella morganii strain MM 190. The maximum level of hemolysis was observed in LB (59%), DMEM supplemented with fetal bovine serum (62%), and urine (53%) under aeration conditions during the exponential growth phase. The presence of 2% urea in the medium suppressed hemolysin synthesis. Moreover, addition of bacterial culture fluid containing hemolysin to a monolayer of T-24 bladder carcinoma and OKP-GS kidney carcinoma cells led to 25 and 42% cell death, respectively. We found that the maximum expression of the hemolysin gene hlyA was observed in 2-h culture in LB medium, which correlated with the hemolytic activity of the bacteria in this medium and indicated the predominance of the short hlyCA transcript in the cells.

[cis-Prenyltransferases of Marchantia polymorpha: Phylogenetic Analysis and Perspectives for Use as Regulators of Antimicrobial Agent Synthesis].

The synthesis of secondary metabolites plays a central role in the survival of plants and their resistance to biotic and abiotic stress. Nevertheless, fundamental and applied studies of plant secondary metabolites, polyisoprenols, remain underdeveloped. The wide distribution of polyisoprenols in plants shows their important role in cellular metabolism. Plant polyisoprenols are synthesized by cis-prenyltransferases (CPTs), the study of which is necessary to understand the synthesis pathways, localization, and functions of plant polyisoprenols. Bryophytes, including the liverwort Marchantia polymorpha, are a unique group of plants with great potential for the study of CPTs. We analyzed the genome of M. polymorpha and identified seven CPT genes, which are homologous to the AtCPT7 and AtCPT3 genes of Arabidopsis thaliana, involved in the synthesis of polyisoprenols. Four individual lines of M. polymorpha plants with mutations in the MpCPT7.37 gene were obtained. It was shown that in three lines the mutation led to a translational frameshift and gene knockout. However, knockout of only the type 7 CPTs had no effect on plant growth and survival. Analysis of the antibacterial activity of mutant plant tissue extracts did not reveal significant changes compared to wild-type tissue extracts, this could be related to a compensatory effect of the activity of other CPTs. These data give us the required base for the further studies of bryophytes CPTs and their products.

Data on the genome analysis of the probiotic strain Bacillus subtilis GM5.

In the present study, we report data on the draft genome sequence of a lipopeptide producing rhizospheric Bacillus subtilis GM5 isolate. The genome consists of 4,271,280 bp with a GC-pair content of 43.3%. A total of 4518 genes including 75 tRNA genes, 3 operons coding for rRNA genes and 56 pseudogenes were annotated. Gene clusters responsible for the biosynthesis of secondary metabolites were validated. Six of the thirty-three clusters identified in the genome code for antimicrobial non-ribosomal peptides synthesis. The Whole Genome Shotgun project of B. subtilis GM5 has been deposited in the NCBI database under the accession number NZ_NKJH00000000 (https://www.ncbi.nlm.nih.gov/nuccore/NZ_NKJH00000000.1).

Structural Characteristics and Catalytic Mechanism of Bacillus ß-Propeller Phytases.

ß-Propeller phytases of Bacillus are unique highly conservative and highly specific enzymes capable of cleaving insoluble phytate compounds. In this review, we analyzed data on the properties of these enzymes, their differences from other phytases, and their unique spatial structures and substrate specificities. We considered influences of different factors on the catalytic activity and thermostability of these enzymes. There are few data on the hydrolysis mechanism of these enzymes, which makes it difficult to analyze their mechanism of action and their final products. We analyzed the available data on hydrolysis by ß-propeller phytases of calcium complexes with myo-inositol hexakisphosphate.

Features of Gene Expression of Bacillus pumilus Metalloendopeptidase.

Features of gene expression of the secreted Bacillus pumilus metalloendopeptidase belonging to the adamalysin/reprolysin family were investigated. In the regulatory region of the gene, we identified hypothetical binding sites for transcription factors CcpA and TnrA. We found that the expression of the metalloendopeptidase gene is controlled by mechanisms of carbon and nitrogen catabolite repression. In experiments involving nitrogen metabolism regulatory protein mutant strains, we found that the control of the metalloendopeptidase gene expression involves proteins of ammonium transport GlnK and AmtB interacting with the TnrA-regulator.

[Effect of Mutations in Extracellular Nuclease on the Characteristics of the Pigmented and Nonpigmented Serratia marcescens Strains].

Comparative characterization of the pigmented and nonpigmented Serratia marcescens strains and their extracellular nuclease mutants was carried out. Biomass accumulation by the mutant strains decreased on average by 20%, while proteolytic activity of the culture liquid was 4-5 times lower than in the case of the wild type strains. The mutants with impaired extracellular nuclease genes exhibited higher sensitivity to reactive oxygen species. Comparative analysis of motility of the strains revealed the highest flagellar activity in the wild type nonpigmented strain, while the cells of its mutant completely lost this feature.

[Peculiarities of Proteus mirabilis extracellular metalloproteinase biosynthesis].

Biosynthesis of metalloproteinase by the Proteus mirabilis 5127-1 strain on different media and the influence of glucose and urea on biosynthesis were studied. It was found that the P. mirabilis 5127-1 bacteria secretes metalloproteinase in the medium in two isoforms (52 and 50 kDa). It was established that proteinase synthesis is completely suppressed during the growth of bacteria on synthetic media, as well as in the presence of LB glucose in the medium. It was demonstrated that addition of urea in the medium results in an increase of the culture productivity in the proteinase synthesis. Maximal culture productivity in the proteinase synthesis was found in the medium with natural urine. During the growth of bacteria on artificial urine, proteinase appeared in the medium only after 12 hours of growth as a single isoform.

[Efflux systems in Serratia marcescens].

A widespread bacterium Serratia marcescens (family Enterobacteriaceae) is an opportunistic and exhibits multiple drug resistance. Active removal of antibiotics and other antimicrobials from pathogen and exhibits multiple drug resistance. Active removal of antibiotics and other antimicrobials from the cells by efflux systems is one of the mechanisms responsible for microbial resistance to these compounds. Among enterobacteria, efflux systems of Escherichia coli and Salmonella enterica var. Typhimurium have been studied most extensively. Few efflux systems that belong to different families have been reported for S. marcescens. In this review, we analyzed available literature about S. marcescens efflux systems and carried out the comparative analysis of the genes encoding the RND type systems in different Serratia species and in other enterobacteria. Bioinformatical analysis of the S. marcescens genome allowed us to identify the previously unknown efflux systems based on their homology with the relevant E. coli genes. Identification of additional efflux systems in S. marcescens genome will promote our understanding of physiology of these bacteria, will detect new molecular mechanisms of resistance and will reveal their resistance potential.

[New metalloendopeptidase of Morganella morganii ZM].

Proteolytic activity which is inhibited in the presence of o-phenanthroline was found in M. morganii ZM. Intracellular proteases of M. morganii ZM unlimited split musculoskeletal actin in contrast to grimelysin. Several proteolitic proteins of M. morganii ZM cells were identified by zymography with gelatin. Metalloproteinase of M. morganii ZM cell lysate was purified by hydrophobic chromatography fractionation. The molecular weight of the protein was 35 kDa.

[Expression optimization of Bacillus pumilus subtilisin-like proteinase].

The characteristics of Bacillus pumilus subtilisin-like protease expression were investigated. Potential binding sites for the transcription factor DegU~P have been identified in the regulatory region of the enzyme gene, which have been optimized for interaction with the regulatory protein. The expression of the extracellular subtilisin-like protease modified gene has been examined. It was established that the modification of one of the sites has resulted in increased expression of the proteinase in 2 times. It is concluded that the optimization of the promoter led to increased expression of subtilisin-like protease.

[Properties of Bacillus pumilus subtilisin like proteinase secreted from recombinant strain on different growth stages].

Bacillus pumilus 3-19 glutamylendopeptidase has been isolated from culture liquid of Bacillus subtilis recombinant strain on different growth stages: growth retardation (early enzyme) and stationary phase (late enzyme). The effect of purified proteinase of different growth stages on insulin beta-chain, protein and oligopeptide substrates has been studied. Comparative study of physicochemical properties of early and late proteinases was carried out. Two protein fractions were different in catalytic characteristics and demonstrated different sensitivity to the presence of metal cations.

[Structural properties and functional importance of metzincin metalloproteinases].

Here wediscuss known properties of metzincin metalloproteinases, their structure, physiological roles in the cell and potential medical uses. We also present results describing a novel extracellular metzincin metalloproteinase from Bacillus pumilus with a unique combination of properties typical for both astacins and adamalysins.

[The novel intracellular phytase of enterobacteriaceae: isolation and properties].

For the first time phytase enzyme was isolated from Pantoea vagans 3.2 strain and was subjected to investigation. The enzyme was purified about 474-fold to apparent homogeneity from the crude extract of the strain, its primary structure was determined and it was concluded that phytase of Pantoea vagans 3.2 belongs to the family of histidine acid phosphatases. It has a molecular mass of about 46 kDa and Km for the hydrolysis of sodium phytate was 0.28 mM. Some physicochemical properties ofphytase were investigated.

[Isolation and characterisation of a new bacillar phytase].

Bacillus ginsengihumi phytase has been firstly isolated and studied from the recombinant Escherichia coli strain cellular lysates. The enzyme was obtained from the cellular lysate, purified till homogeneous condition, primary structure was determined. It's concluded that phytase relates to beta-propeller class of phosphatases. The molecular weight of the protein was 41 kDa, pI was 4.8. Some physical and chemical properties of the enzyme were studied.

[The novel Adams-like microbial metalloendopeptidase].

Heterologous gene expression of extracellular minor metalloendopeptidase of Bacillus pumilus 3-19 in protease-deficient B. subtilis strain has been studied. The fraction of enzyme in total pool of B. pumilus 3-19 secreted proteases composes less than 8%. The enzyme was isolated from culture liquid of recombinant strain, its primary structure was determined, physicochemical properties were investigated. It was concluded that secreted metallo endopeptidase of B. pumilus 3-19 represents the first prokaryotic homolog of eukaryotic adamalysin/reprolysin protein family.