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Biotechnology Reports (Amsterdam,... Sep 2020Alginate is a group of water-soluble linear polysaccharides comprising of variable units of α-l-guluronic and β-d-mannuronic acid. The alginates are in high demand in...
Alginate is a group of water-soluble linear polysaccharides comprising of variable units of α-l-guluronic and β-d-mannuronic acid. The alginates are in high demand in biomedical, pharmaceutical and bioengineering applications. In the present study, we have isolated a strain of that has potential alginate synthesis. The biochemical and physiochemical characteristic including Carbazole assay, DSC, FTIR and H NMR were confirmed the alginate synthesis efficacy by . Evaluation of alginate for the removal of heavy metals such as Chromium, Cobalt and Lead showed that it effectively adsorbs heavy metals. Further analysis of gelling ability and cytotoxicity evaluation revealed that the alginate can be reconstituted as hydrogel and scaffold. Overall, our findings suggest that the strain may be used to produce alginate at commercial level that has the potential bioremediation and biomedical applications.
PubMed: 32874945
DOI: 10.1016/j.btre.2020.e00517 -
Bioresources and Bioprocessing May 2022Prokaryotic Argonaute (pAgo) proteins are well-known oligonucleotide-directed endonucleases, which contain a conserved PIWI domain required for endonuclease activity....
BACKGROUND
Prokaryotic Argonaute (pAgo) proteins are well-known oligonucleotide-directed endonucleases, which contain a conserved PIWI domain required for endonuclease activity. Distantly related to pAgos, PIWI-RE family, which is defined as PIWI with conserved R and E residues, has been suggested to exhibit divergent activities. The distinctive biochemical properties and physiological functions of PIWI-RE family members need to be elucidated to explore their applications in gene editing.
RESULTS
Here, we describe the catalytic performance and cellular functions of a PIWI-RE family protein from Pseudomonas stutzeri (PsPIWI-RE). Structural modelling suggests that the protein possesses a PIWI structure similar to that of pAgo, but with different PAZ-like and N-terminal domains. Unlike previously reported pAgos, recombinant PsPIWI-RE acts as an RNA-guided DNA nuclease, as well as a DNA-guided RNA nuclease. It cleaves single-stranded DNA at temperatures ranging from 20 to 65 °C, with an optimum temperature of 45 °C. Mutation at D525 or D610 significantly reduced its endonuclease activity, confirming that both residues are key for catalysis. Comparing with wild-type, mutant with PIWI-RE knockout is more sensitive to ciprofloxacin as DNA replication inhibitor, suggesting PIWI-RE may potentially be involved in DNA replication.
CONCLUSION
Our study provides the first insights into the programmable nuclease activity and biological function of the unknown PIWI-RE family of proteins, emphasizing their important role in vivo and potential application in genomic DNA modification.
PubMed: 38647609
DOI: 10.1186/s40643-022-00539-x -
Archives of Microbiology Jul 2022Textile wastewater contains dyes mixed with other contaminants in various concentrations. Bacteria-mediated decolorization and degradation of azo dyes have achieved...
Textile wastewater contains dyes mixed with other contaminants in various concentrations. Bacteria-mediated decolorization and degradation of azo dyes have achieved momentum as a method of treatment attributed to their inexpensive, eco-friendly, and application to a wide range of azo dyes. However, a single species of bacteria is inefficient in decolorizing diverse groups of dyes which is one of the most significant challenges for environmental technologists working in bioremediation. In the present study, an aerobic bacterial consortium AUJ consisting of six different bacterial strains (Pseudomonas stutzeri AK1, Pseudomonas stutzeri AK2, Pseudomonas stutzeri AK3, Bacillus spp. AK4, Pseudomonas stutzeri AK5, and Pseudomonas stutzeri AK6) removed the individual azo dyes in the 24-94% range when used in more than 200 ppm concentration within 72-96 h. In addition, the consortium was able to decolorize 52.19% mixed dyes (100 ppm) and 44.55% Acid blue 113 when used at a concentration as high as 1100 ppm within 96 h. Optimization of various nutritional and environmental parameters revealed that glucose and yeast extract were the preferred carbon and nitrogen source, respectively, and analysis of treated dye products using high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FTIR), and gas chromatography-mass spectrometry (GC-MS) confirmed the breakdown of dye. In all, we present a bacterial consortium with a good ability of dye decolorization that can be used for degrading a wide variety of azo dyes.
Topics: Azo Compounds; Bacteria; Biodegradation, Environmental; Coloring Agents; Wastewater
PubMed: 35867172
DOI: 10.1007/s00203-022-03108-0 -
PloS One 2020High concentrations of metals in the environment alter bacterial diversity, selecting resistant and tolerant species. The study evaluated the selection of a potential...
High concentrations of metals in the environment alter bacterial diversity, selecting resistant and tolerant species. The study evaluated the selection of a potential bacterial strain from Sepetiba Bay-Rio de Janeiro, Brazil marine sediments to remove Cu and Pb. The bacterial strain isolated from the sediments was used in three different bioassays: (1) Cu at concentrations of 0 (control), 6 and 50 μg.mL-1; (2) Pb at concentrations of 0 (control), 6 and 50 μg.mL-1; (3) Cu + Pb in concentrations of 3 μg.mL-1 Cu + 3 μg.mL-1 Pb (6 μg.mL-1) and 25 μg.mL-1 Cu + 25 μg.mL-1 Pb (50 μg.mL-1). The number of cells and the enzymatic activities of dehydrogenases and esterases were quantified. Results of taxonomic identification indicated the selection of the Pseudomonas stutzeri W228 strain, showing a greater degree of similarity (±73%) with the database used. There was no significant variation in the number of cells, 108 cells.mL-1, which represents a high biomass production in the presence of stressors. However, we observed a reduction in dehydrogenase activity at all tested concentrations of Cu, Pb and Cu + Pb. The activity of esterase increased, indicating a higher energy demand to complete the bacterial life cycle. The study showed significant results for the absorption of Pb by the extracellular polymeric substances (EPS) and the efflux of Cu. The capacity of Pb absorption by EPS can be considered a resistance mechanism, as well as the efflux of Cu, so that the available EPS sites could be occupied by the most toxic ions demonstrating that Pseudomonas stutzeri is resistant to Pb and Cu.
Topics: Bacterial Proteins; Bacteriological Techniques; Biodegradation, Environmental; Biomass; Brazil; Copper; Esterases; Extracellular Polymeric Substance Matrix; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Geologic Sediments; Lead; Oxidoreductases; Pseudomonas stutzeri; Water Pollutants, Chemical
PubMed: 33104697
DOI: 10.1371/journal.pone.0240486 -
International Journal of Nanomedicine 2020Selenium nanoparticles (SeNP) have several applications in the field of biotechnology, including their use as anti-cancer drugs. The purpose of the present study is to...
PURPOSE
Selenium nanoparticles (SeNP) have several applications in the field of biotechnology, including their use as anti-cancer drugs. The purpose of the present study is to analyze the efficacy of green synthesis on the preparation of SeNP and its effect on their anti-cancer properties.
METHODS
A bacterial strain isolated from a freshwater source was shown to efficiently synthesize SeNP with potential therapeutic properties. The quality and stability of the NP were studied by scanning electron microscopy, X-ray diffraction, zeta-potential and FTIR analysis. A cost-effective medium formulation from biowaste having 6% banana peel extract enriched with 0.25 mM tryptophan was used to synthesize the NP. The NP after optimization was used to analyze their anti-tumor and anti-angiogenic activity. For this purpose, first, the cytotoxicity of the NP against cancer cells was analyzed by MTT assay and then chorioallantoic membrane assay was performed to assess anti-angiogenic activity. Further, cell migration assay and clonogenic inhibition assay were performed to test the anti-tumor properties of SeNP. To assess the cytotoxicity of SeNP on healthy RBC, hemolysis assay was performed.
RESULTS
The strain identified as (MH191156) produced phenazine carboxylic acid, which aids the conversion of Se oxyanions to reduced NP state, resulting in particles in the size range of 75 nm to 200 nm with improved stability and quality of SeNP, as observed by zeta (ξ) potential of the particles which was found to be -46.2 mV. Cytotoxicity of the SeNP was observed even at low concentrations such as 5 µg/mL against cervical cancer cell line, ie, HeLa cells. Further, neovascularization was inhibited by upto 30 % in CAMs of eggs coinoculated with SeNp when compared with untreated controls, indicating significant anti-angiogenic activity of SeNP. The NP also inhibited the invasiveness of HeLa cells as observed by decreased cell migration and clonogenic proliferation. These observations indicate significant anti-tumor and anti-angiogenic activity of the SeNP in cervical cancer cells.
CONCLUSION
(MH191156) is an efficient source of Se NP production with potential anti-angiogenic and anti-tumor properties, particularly against cervical cancer cells.
Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Cell Death; Cell Movement; Cell Proliferation; Cell Survival; Chick Embryo; Costs and Cost Analysis; Female; HeLa Cells; Hemolysis; Humans; Metal Nanoparticles; Phenazines; Pseudomonas stutzeri; Reproducibility of Results; Selenium; Spectroscopy, Fourier Transform Infrared; Static Electricity; Uterine Cervical Neoplasms; X-Ray Diffraction
PubMed: 32606692
DOI: 10.2147/IJN.S247426 -
The Science of the Total Environment Aug 2020Functional durability of bio-augmented microbes in contaminated fields remains a major challenge in bioremediation. In the present study, various immobilization...
Functional durability of bio-augmented microbes in contaminated fields remains a major challenge in bioremediation. In the present study, various immobilization materials and compositional combinations were designed and compared to enhance the functional durability of Pseudomonas stutzeri sp. Y2 for degradation of simazine, one of the most used herbicides, in industrial wastewater and maize fields. Among four combinations of materials tested, the optimal combination obtained from the orthogonal array trials was 14% polyvinyl alcohol (PVA), 1-3% sodium alginate (SA), 2% activated carbon (AC), and 1-2% Y2 cells (PSC-Y2), which yielded 1.7 fold faster degradation of simazine at 50 mg L than that by free Y2 cells in the industrial wastewater. The degradation half-lives (DT) of simazine (10 mg L) by free Y2 cells and PSC-Y2 was 1.1 d and 5.3 d in laboratory soil, respectively. The DT of simazine by PSC-Y2 at the recommended and double dosages of simazine (0.45 and 0.9 g ai·m) was 17.2 d and 12.4 d in the maize fields, respectively, in comparison with 23 d and 17.4 d by free Y2 cells. In addition, the PSC-Y2 degraded 100% of atrazine and terbuthylazine, and 96% of propazine at an initial concentration of 50 mg L each in 4 days. This study provides an immobilization strategy to stabilize bacteria and prolong bacterial functions to treat s-triazine herbicides contaminated water and soil.
Topics: Atrazine; Herbicides; Pseudomonas stutzeri; Simazine; Triazines; Wastewater; Zea mays
PubMed: 32388161
DOI: 10.1016/j.scitotenv.2020.139183 -
AMB Express May 2022Psychrotrophic Pseudomonas is one of the significant microbes that lead to putrefaction in chilled meat. One of the biggest problems in the detection of Pseudomonas is...
Psychrotrophic Pseudomonas is one of the significant microbes that lead to putrefaction in chilled meat. One of the biggest problems in the detection of Pseudomonas is that several species are seemingly identical. Currently, antibiotic resistance is one of the most significant challenges facing the world's health and food security. Therefore, this study was designed to apply an accurate technique for eliminating the identification discrepancy of Pseudomonas species and to study their resistance against various antimicrobials. A total of 320 chicken meat specimens were cultivated, and the isolated bacteria' were phenotypically recognized. Protein analysis was carried out for cultured isolates via Microflex LT. The resistance of Pseudomonas isolates was recorded through Vitek® 2 AST-GN83 cards. Overall, 69 samples were identified as Pseudomonas spp. and included 18 Pseudomonas lundensis (P. lundensis), 16 Pseudomonas fragi (P. fragi), 13 Pseudomonas oryzihabitans (P. oryzihabitans), 10 Pseudomonas stutzeri (P. stutzeri), 5 Pseudomonas fluorescens (P. fluorescens), 4 Pseudomonas putida (P. putida), and 3 Pseudomonas aeruginosa (P. aeruginosa) isolates. Microflex LT identified all Pseudomonas isolates (100%) correctly with a score value ≥ 2.00. PCA positively discriminated the identified isolates into various groups. The antimicrobial resistance levels against Pseudomonas isolates were 81.16% for nitrofurantoin, 71% for ampicillin and ampicillin/sulbactam, 65.22% for cefuroxime and ceftriaxone, 55% for aztreonam, and 49.28% for ciprofloxacin. The susceptibilities were 100% for cefotaxime, 98.55% for ceftazidime, 94.20% for each piperacillin/tazobactam and cefepime, 91.3% for cefazolin. In conclusion, chicken meat was found to be contaminated with different Pseudomonas spp., with high incidence rates of P. lundensis. Microflex LT is a potent tool for distinguishing Pseudomonads at the species level.
PubMed: 35532863
DOI: 10.1186/s13568-022-01390-1 -
Journal of Infection and Public Health 2019We report a case of Pseudomonas stutzeri endocarditis in Lebanon. The patient had a recent history of prosthetic aortic valve replacement and presented to the emergency... (Review)
Review
We report a case of Pseudomonas stutzeri endocarditis in Lebanon. The patient had a recent history of prosthetic aortic valve replacement and presented to the emergency department with fever and chills. Transesophageal echocardiography confirmed the presence of a vegetation on the prosthetic valve and blood cultures yielded P. stutzeri. The patient was treated with surgery and antibiotics but deteriorated and passed away four days after admission. To our knowledge, this is the fifth case of P. stutzeri endocarditis reported in the literature, and the first case with early presentation and mortality.
Topics: Aged; Anti-Bacterial Agents; Aortic Valve; Ceftazidime; Chills; Diagnosis, Differential; Echocardiography, Transesophageal; Endocarditis, Bacterial; Heart Valve Prosthesis; Heart Valve Prosthesis Implantation; Humans; Infusions, Intravenous; Male; Postoperative Complications; Prosthesis-Related Infections; Pseudomonas stutzeri
PubMed: 30049610
DOI: 10.1016/j.jiph.2018.07.004 -
Analytical Chemistry Aug 2023Small proteins of around 50 aa in length have been largely overlooked in genetic and biochemical assays due to the inherent challenges with detecting and characterizing...
Small proteins of around 50 aa in length have been largely overlooked in genetic and biochemical assays due to the inherent challenges with detecting and characterizing them. Recent discoveries of their critical roles in many biological processes have led to an increased recognition of the importance of small proteins for basic research and as potential new drug targets. One example is CcoM, a 36 aa subunit of the -type oxidase that plays an essential role in adaptation to oxygen-limited conditions in , a model for the clinically relevant, opportunistic pathogen . However, as no comprehensive data were available in , we devised an integrated, generic approach to study small proteins more systematically. Using the first complete genome as basis, we conducted bottom-up proteomics analyses and established a digest-free, direct-sequencing proteomics approach to study cells grown under aerobic and oxygen-limiting conditions. Finally, we also applied a proteogenomics pipeline to identify missed protein-coding genes. Overall, we identified 2921 known and 29 novel proteins, many of which were differentially regulated. Among 176 small proteins 16 were novel. Direct sequencing, featuring a specialized precursor acquisition scheme, exhibited advantages in the detection of small proteins with higher (up to 100%) sequence coverage and more spectral counts, including sequences with high proline content. Three novel small proteins, uniquely identified by direct sequencing and not conserved beyond , were predicted to form an operon with a conserved protein and may represent genes. These data demonstrate the power of this combined approach to study small proteins in and show its potential for other prokaryotes.
Topics: Pseudomonas stutzeri; Proteomics; Proteogenomics; Pseudomonas aeruginosa; Oxygen
PubMed: 37535005
DOI: 10.1021/acs.analchem.3c00676 -
Turkish Journal of Pharmaceutical... Feb 2021Tamoxifen (TAM), which is used for treating breast cancer, has exhibited another important function as an antimicrobial agent. The objective of this study is to...
OBJECTIVES
Tamoxifen (TAM), which is used for treating breast cancer, has exhibited another important function as an antimicrobial agent. The objective of this study is to investigate the antibacterial action of TAM against the bacteria present in the human oral cavity.
MATERIALS AND METHODS
The bacteria present in the human oral cavity were isolated from healthy individuals. Different concentrations of TAM were tested against the isolated bacteria. Additionally, bactericidal and bacteriostatic effects of TAM were also determined.
RESULTS
Out of 23 isolated bacteria, a greater number of Gram-positive bacteria were highly susceptible to the low concentrations of TAM than Gram-negative bacteria. , which is Gram-positive bacterium, and , which is Gram-negative bacterium, needed a high minimum inhibitory concentration value of TAM (2.5 mg/mL) to be inhibited by TAM's bacteriostatic action. Resistance to TAM was also observed in three strains of Gram-positive and four strains of Gram-negative bacteria.
CONCLUSION
TAM has shown a potential antibacterial effect against the bacteria present in the oral cavity, especially against Gram-positive bacteria. This effect is mostly bacteriostatic. This study also found bacterial resistance toward TAM.
PubMed: 33634670
DOI: 10.4274/tjps.galenos.2019.23500