Moderately haloalkaliphilic aerobic methylobacteria

Aerobic methylobacteria utilizing oxidized and substituted methane derivatives as carbon and energy sources are widespread in nature and involved in the global carbon cycle, being a unique biofilter on the path of these C1 compounds from different ecosystems to the atmosphere. New data on the biological features of moderately halophilic, neutrophilic, and alkaliphilic methylobacteria isolated from biotopes with higher osmolarity (seas, saline and soda lakes, saline soils, and deteriorating marble) are reviewed. Particular attention is paid to the latest advances in the study of the mechanisms of osmoadaptation of aerobic moderately haloalkaliphilic methylobacteria: formation of osmolytes, in particular, molecular and genetic aspects of biosynthesis of the universal bioprotectant ectoine. The prospects for further studies of the physiological and biochemical principles of haloalkalophily and for the application of haloalkaliphilic aerobic methylobacteria in biosynthesis and biodegradation are discussed.     

Genes involved in the regulation of vascular homeostasis determine renal survival rate in patients with chronic glomerulonephritis

Chronic glomerulonephritis (CGN) is one of the most severe kidney diseases. Genes of vascular reactivity are thought to play an important role in development and progression of CGN. In this study, we analyzed association of genes of vascular homeostasis with hypertension and renal survival of CGN patients. The study sample included 238 patients with CGN and 304 healthy subjects of population control. Ten polymorphisms of ten genes of vascular homeostasis were genotyped through polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) analysis and TaqMan assays. Association of the genotypes with renal survival was analyzed by the Kaplan–Meier estimator. Genotypes 311SC and 311SS of the PON2 gene, (− 1166)AC and (− 1166)CC of the AGTR1 gene, (+ 46)AA of the ADRB2 gene, and 198KK and 198KN of the EDN1 gene were associated with decreased rate of renal survival of the patients. Polymorphisms S311C PON2, (− 1166)A/C AGTR1, (+ 46)G/A ADRB2, and K198N EDN1 were associated with the accelerated decline in kidney function in the CGN patients.     

Range expansion of rotan <Emphasis Type="Italic">Perccottus glenii</Emphasis>, sunbleak <Emphasis Type="Italic">Leucaspius delineatus</Emphasis>, and bleak <Emphasis Type="Italic">Alburnus alburnus</Emphasis> in the Ob River basin

The ranges of three alien fish species (rotan Perccottus glenii, sunbleak Leucaspius delineatus, and bleak Alburnus alburnus) in the Ob River basin, West Siberia, have expanded. Our data prove the expansion of rotan and sunbleak ranges to the north. In particular, for the first time, populations of rotan are reported from the lower section of the Ob. We also present new data about range expansion of the studied fish species southward. All three species have probably reached the southern limits of their possible distribution in the Upper Ob system, entering the potamon/rhithron transition zone, because the species under study prefer lowland waters. The spatio-temporal dynamics of their expansion reveals the role of the Ob River as the main invasion corridor, enhancing their distribution mostly in meridional direction. In the system of the Upper Ob, rotan and sunbleak were found by us in isolated ponds and lakes, as well as in floodplain systems. This is in agreement with the concept of two invasion vectors: the translocation of fish by humans between water bodies and self-distribution along the river courses.     

Q-band chromosome polymorphism in couples with reproductive disorders

A comparative analysis of Q-band polymorphisms was conducted in phenotypically normal individuals and couples with repeated spontaneous abortions. In the group of pathology reduction of the following phenomena was found: frequencies of brilliant fluorescence in bands 4p11q11, 14p11, 14p13, 15p11, 21p13; the mean numbers of the brilliant fluorescent segments per cell per individual; frequencies of extremely large ("marker") Q-bands. All these findings coincided with our earlier published data on reduction of the C-heterochromatin amount in the same group of pathology.     

Use of protein-free potato juice concentrates in the production of tetracycline]

The study on the use of potato juice concentrate in tetracycline production showed that the tetracycline-producing strain 3-NK of Actinomyces aureofaciens grown on seed and fermentation media containing 2--2.5 and 0.35--0.45 per cent (by dry substance) of the concentrate produced the same antibiotic yield as that on the media with corn steep liquor.     

Prevalence of Helicobacter pylori CagA-producing strains within families

During prophylactic examination of blood sera taken from the members of 59 families by the enzyme immunoassay, antibodies to H. pylori and CagA protein were determined. As shown in this study, the children of non-infected mothers proved to be infected in 6.3% of cases and the children of infected mothers, in 72.1% of cases (p < 0.001). The children of non-infected fathers were H. pylori-positive in 71.4% and those of infected fathers, in 58.4% of cases. The CagA status was found to coincide in mothers and their children (p = 0.01), but not in fathers and their children. These data indicate that children acquire H. pylori infection from the members of their family, mainly from their mothers.     

The genes and enzymes of sucrose metabolism in moderately thermophilic methanotroph Methylocaldum szegediense O12

Extremophiles - Four enzymes involved in sucrose metabolism: sucrose phosphate synthase (Sps), sucrose phosphate phosphatase (Spp), sucrose synthase (Sus) and fructokinase (FruK), were obtained as...     

Homo- and heterologous reporter proteins for evaluation of promoter activity in <Emphasis Type="Italic">Methylomicrobium alcaliphilum</Emphasis> 20Z

A number of vectors were constructed based on the plasmid from the broad range of pMHA200 hosts. Also, the expression of some key genes of the haloalkalitolerant methanotroph Methylomicrobium alcaliphilum 20Z was studied. The activities of the promoter regions of genes for hexulose phosphate synthase, glutamine synthetase, and glucokinase, as well as the promoter of the ectABC-ask operon, which encodes enzymes for osmoprotectant ectoine biosynthesis, were evaluated with the use of the gfp gene; the evaluation was proven to be ineffective. Conversely, glucokinase and a heterologous enzyme of chloramphenicol acetyltransferase were useful for the evaluation of promoter activity. In M. alcaliphilum 20Z cells, the expression level of chloramphenicol acetyltransferase transcribed from the methanol dehydrogenase promoter was higher as compared with that of glucokinase. This seems to be due to a regulatory mechanism for homologous protein expression. The introduction of a synthetic nucleotide sequence forming the secondary structure in the 5′ untranslated region of the glucokinase mRNA resulted in an increase of this enzyme level. This is the first attempt to use M. alcaliphilum 20Z for homo- and heterologous protein expression.     

Alteration of cofactor specificity of the acrylyl-CoA reductase from Escherichia coli

Objectives

Alteration of the cofactor specificity of acrylyl-CoA reductase (AcuI) catalyzing the NAD(P)H-dependent reduction of acrylyl-CoA to propionyl-CoA is often desirable for designing of artificial metabolic pathways of various appointments.

Results

Several variants of AcuIs from Escherichia coli K-12 with multiple amino acid substitutions to alter the cofactor preference were obtained by site directed mutagenesis and the modified enzymes as His₆-tagged proteins were characterized. The simultaneous substitutions of arginine-180, arginine-198 and serine-178 residues by alanine in the enzyme pocket sequence as well as other amino acid changes decreased both NADPH- and NADH-dependent activities in comparison to the wild-type enzyme. The replacement of serine-156 by glutamic acid decreased NADPH-dependent activity at least 7000-fold but NADH-dependent activity only by threefold. The replacement of serine-156 by aspartic acid decreased NADPH-dependent activity 70-fold with fair preservation of activity and specificity to NADH.

Conclusions

These results demonstrated a relevance of Asp156 in the interaction of AcuI from E. coli K-12 with NADH as a coenzyme. These findings may provide reference information for shifting coenzyme specificity of acrylyl-CoA reductases.