Lytic transglycosylases: concinnity in concision of the bacterial cell wall

The lytic transglycosylases (LTs) are bacterial enzymes that catalyze the non-hydrolytic cleavage of the peptidoglycan structures of the bacterial cell wall. They are not catalysts of glycan synthesis as might be surmised from their name. Notwithstanding the seemingly mundane reaction catalyzed by the LTs, their lytic reactions serve bacteria for a series of astonishingly diverse purposes. These purposes include cell-wall synthesis, remodeling, and degradation; for the detection of cell-wall-acting antibiotics; for the expression of the mechanism of cell-wall-acting antibiotics; for the insertion of secretion systems and flagellar assemblies into the cell wall; as a virulence mechanism during infection by certain Gram-negative bacteria; and in the sporulation and germination of Gram-positive spores. Significant advances in the mechanistic understanding of each of these processes have coincided with the successive discovery of new LTs structures. In this review, we provide a systematic perspective on what is known on the structure–function correlations for the LTs, while simultaneously identifying numerous opportunities for the future study of these enigmatic enzymes.     

Conventional and Molecular Identification of Bacteria with Magnesite Enrichment Potential from Local Quarries in Erzurum

In this study, the bacteria having ore enrichment potential were isolated from three different magnesite quarries located in Erzurum-Askale borderlines. The obtained isolates were identified and characterized according to the conventional (morphological, physiological and biochemical tests) and molecular techniques (fatty acid methyl ester profiles (FAME), BOX PCR and 16S rDNA). According to sequence analysis, they were determined as Exiguobacterium aurantiacum (4), Exiguobacterium sibiricum (2), Bacillus sp. (2), Staphylococcus epidermidis (2), Staphylococcus haemolyticus (1), Shewanella baltica (1) and Klebsiella oxytoca (1), respectively.     

Secondary productivity (3H-Leucine and 3H-Thymidine incorporation), abundance and biomass of the epiphytic bacteria attached to detritus of Typha domingensis pers. in a tropical coastal lagoon

Senescent, naturally dried leaves of Typha domingensis were incubated inthe littoral region of a coastal lagoon and epiphytic bacterial volume,abundance, biomass and secondary productivity were measured during 127 daysof decomposition. The peak of cell abundance was registered at t =127 days when expressed per leaf surface area (10.07×10⁷cells cm^-2; 7.26 µgC cm^-2), and at t= 26 days when expressed per biofilm dry mass (38.10 ×10⁷ cells (mgDM biofilm)^-1, 30.52 µgC(mgDM biofilm)^-1). The highest values of bacterial biovolumesand lower turnover time were usually obtained in the beginning of thecolonization. Leu:Tdr ratios were also higher in the beginning of thecolonization, when bacterial community presented unbalanced metabolism.Consequently, the highest discrepancies between the bacterial secondaryproduction estimated by leu and Tdr incorporation were observed in the first2 days of decomposition. On average, the bacterial secondary productivityestimated by leu incorporation was 2.1 times higher than the valuesestimated by Tdr incorporation when the empirical factor for Tdr wasobtained from the relationship between Tdr and biomass increment. Thisdifference increased to 4.2 when the empirical factor was obtained from therelationship between Tdr and cell numbers increment. An average of bothmethods (0.0037 to 0.1397 µgC cm^-2 h^-1)produced results that fall within the range reported in the literature forepiphytic bacteria of freshwater ecosystems.     

A method to study zhizosphere processes in thin soil layers of different proximity to roots

Frankia is the diverse bacterial genus that fixes nitrogen within root nodules of actinorhizal trees and shrubs. Systematic and ecological studies of Frankia have been hindered by the lack of morphological, biochemical, or other markers to readily distinguish strains. Recently, nucleotide sequence of 16 S RNA from the small ribosomal subunit has been used to classify and identify a variety of microorganisms. We report nucleotide sequences from portions of the 16 S ribosomal RNA from Frankia strains AcnI1 isolated from Alnus viridis ssp. crispa (Ait.) Turrill and PtI1 isolated from Purshia tridentata (Pursh) DC. The number of nucleotide base substitutions and gaps we find more than doubles the previously reported sequence diversity for the same variable regions within other strains of Frankia.     

Polyphasic characterization of Delftia acidovorans ESM-1, a facultative methylotrophic bacterium isolated from rhizosphere of Eruca sativa

In this study, one bacterial strain, ESM-1, was isolated from rhizosphere of Eruca sativa, growing in Al Hofouf, Saudia Arabia, after enrichment with methanol as a sole carbon and energy source in a batch culture. ESM-1 was characterized by a polyphasic approach. The strain was identified as Delftia acidovorans at similarity level of 99.9% of the 16S rRNA gene sequences. Results of the Biolog Gen III MicroPlate test system showed that strain ESM-1 reacted positively to 47 (50%) including the one-carbon compound formic acid, and partially positive to 6 (∼6.4%) out of the 94 different the traits examined. The total cellular fatty acids composition of the strain ESM-1 was (C_16:1ω7c/C_16:1ω6c) and C_16:0) and matched that of Delftia acidovorans at a similarity index of 0.9, providing a robustness to the ESM-1 identification. Furthermore, ESM-1 displayed a complex polar lipid profile consisting of phosphatidylethanolamine, phosphatidylglycerol, glycolipid, aminolipid, in addition to uncharacterized lipids. The DNA G+C content of the strain was 66.6 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain ESM1-1 was clearly clustered within the Delftia clade and constructed a monophyletic subcluster with Delftia acidovorans NBRC14950. The results addressed that ESM-1 is a facultative methylotrophic bacterium indigenous to Al Hofouf region and opens the door for potential biotechnological applications (e.g., bioremediation) of this strain, in future. Additionally, these findings assure that the total cellular fatty acid analysis and 16S rRNA gene are reliable tool for bacterial characterization and identification.     

Purification and characterization of β-galactosidase from probiotic Pediococcus acidilactici and its use in milk lactose hydrolysis and galactooligosaccharide synthesis

β-galactosidase is a commercially important enzyme that was purified from probiotic Pediococcus acidilactici. The enzyme was extracted from cells using sonication and subsequently purified using ammonium sulphate fractionation and successive chromatographies on Sephadex G-100 and Q-Sepharose. The enzyme was purified 3.06-fold up to electrophoretic homogeneity with specific activity of 0.883 U/mg and yield of 28.26%. Molecular mass of β-galactosidase as estimated by SDS-PAGE and MALDI-TOF was 39.07 kDa. The enzyme is a heterodimer with subunit mass of 15.55 and 19.58 kDa. The purified enzyme was optimally active at pH 6.0 and stable in a pH range of 5.8–7.0 with more than 97% activity. Purified β-galactosidase was optimally active at 50 °C. Kinetic parameters K_m and V_max for purified enzyme were 400 µM and 1.22 × 10⁻¹ U respectively. Its inactivation by PMSF confirmed the presence of serine at the active site. The metal ions had different effects on enzyme. Ca²⁺, Mg²⁺ and Mn²⁺ slightly activated the enzyme whereas NH₄⁺, Co²⁺ and Fe³⁺ slightly decreased the enzyme activity. Thermodynamic parameters were calculated that suggested that β-galactosidase is less stable at higher temperature (60 °C). Purified enzyme effectively hydrolysed milk lactose with lactose hydrolysing rate of 0.047 min⁻¹ and t_1/2 of 14.74 min. This is better than other studied β-galactosidases. Both sonicated Pediococcus acidilactici cells and purified β-galactosidase synthesized galactooligosaccharides (GOSs) as studied by TLC at 30% and 50% of lactose concentration at 47.5 °C. These findings indicate the use of β-galactosidase from probiotic bacteria for producing delactosed milk for lactose intolerant population and prebiotic synthesis. pH and temperature optima and its activation by Ca²⁺ shows that it is suitable for milk processing.     

兰州唐古特白刺根际微生物分离鉴定

【目的】从兰州唐古特白刺根际分离得到对植物有潜在促生效果的功能微生物，为研发相关菌种制剂的研究奠定基础。【方法】通过平板划线法从其根际分离纯化出6株细菌，并对菌株进行形态特征观察、革兰氏染色等一系列生理生化试验。用藜麦检测各菌株的促生功能，并对具有优良促生作用的1个菌株16S rRNA基因进行分子鉴定及基因草图绘制。【结果】根据生化鉴定结果，6株细菌分别属于不动杆菌属(Acinetobacter)、土壤杆菌属(Agrobacterium)、类芽孢杆菌属(Paenibacillus)、芽孢杆菌属(Bacillus)、鞘氨醇单胞菌属(Sphingomonas)和假单胞菌属(Pseudomonas)。其中，16S rRNA基因鉴定BC4属于肠杆菌属(Enterobacter)，具有较好的促生效果。【结论】BC4具有较好的促生效果，为兰州唐古特白刺菌种资源的开发和利用提供了一定的理论依据。     

Correlation between the parameters of contingent negative variation and characteristics of variational pulsometry in Parkinsonian patients

In patients suffering from Parkinson's disease (PD), we analyzed correlations between the parameters of contingent negative variation (CNV) and data of variational pulsometry (according to the measurements of R-R ECG intervals). Studies were carried out on 35 patients (group PD), 49 to 74 years old, with the stage of disease of 1.5 to 3.0 according to the Hoehn-Yahr international classification. In the course of CNV recording (i.e., in the state of a certain functional loading), we observed significant negative correlations between the integral magnitude (area) of this potential and indices of variational pulsometry (RMSSD, SDNN, C. var, and HF) that characterize the intensity of parasympathetic (respiratory) influences on the cardiovascular system. In the control group, such correlations were absent. We found significant correlations between the autonomic balance, CNV magnitude, and stage of PD reflecting the level of generalization of the pathological process. In the subgroup of patients with the PD stage 1.5 to 2.0, significant changes in the mean values of indices of parasympathetic influences during recording of the CNV were not observed, while in another subgroup (the PD stage 2.5 to 3.0), these values increased significantly (P < 0.05 and P < 0.01). If the estimates of the PD stage were low, the CNV area demonstrated greater values (P < 0.01). The disturbance of coordination of muscle-to-muscle interactions in the PD group is, probably, an important factor responsible for parasympathetic dysregulation and suppression of the CNV generation. We found positive correlation between the intensity of parasympathetic influences in the course of CNV recording and the level of postural disorders (r = 0.37, P < 0.05). On the contrary, the CNV magnitude demonstrated a negative correlation with the intensity of these disorders (r = −0.36, P < 0.05), as well as with the level of postural instability (r = −0.55, P < 0.001). We hypothesize that alterations of the autonomic balance and the activity of those cerebral structures, which are responsible for the motor readiness, result, to a significant extent, from weakening of the activity of the noradrenergic system due to degenerative processes developing in cells of the locus coeruleus. The impairment of the latter structure, together with degeneration of neurons of the substantia nigra and a decrease in the level of nigro-striatal dopamine, underlies the pathomorphological pattern of PD. Neirofiziologiya/Neurophysiology, Vol. 40, No. 3, pp. 242–253, May–June, 2008.     

Adsorption of DNA by Bacteria and Their Composites with Minerals

Previous studies revealed the thermodynamic properties of DNA adsorption on pure minerals or biomasses; however, there has been little attempt to develop such studies on bacteria–mineral composites. Equilibrium adsorption experiments, attenuated total reflectance Fourier transform infrared spectroscopy, and isothermal titration calorimetry were employed to investigate the adsorption of DNA by Bacillus subtilis, Pseudomonas putida, and their composites with minerals. Similar capacity and affinity were observed for DNA adsorption on two bacterial cells. However, different patterns were found in the adsorption of DNA by bacteria–mineral composites. The Gram-positive bacterium B. subtilis enhanced the adsorption of DNA on its mineral composites compared with their individual components, while the composites of Gram-negative bacterial cells with kaolinite and goethite bound lower amounts of DNA than the predicted values. The thermodynamic parameters and the Fourier transform infrared spectra showed that van der Waals force and hydrogen bonding are responsible for the DNA adsorption on B. subtilis–minerals and P. putida–kaolinite. By contrast, the entropy increases of excluded water rearrangement and dehydration effect play key roles in the interaction between DNA and P. putida–montmorillonite/goethite composites.