RESUMO
Iron sequestration through ferritin forms a major part of innate immune response in molluscs and detailed understanding of ferritin gene and its functions can be directly applied in infection and disease management studies. Accordingly, identification and detailed molecular characterization of a ferritin subunit gene from a commercially significant marine mussel Perna viridis was targeted. Molecular screening using degenerate primers in total mantle RNA resulted in the amplification of a novel ferritin gene fragment having <87% identity to the reported ferritin gene sequences. Rapid amplification of cDNA ends-PCR was followed to generate complete cDNA sequence of P.viridis ferritin (PvFer). The complete cDNA was found to be 798 bp, containing an open reading frame of 522 bp, 5' untranslated region (UTR) of 112 bp and 3' UTR of 165 bp. The 5' UTR and 3' UTR were shown to contain an iron response element (IRE) and a polyadenylation signal (767AATAAA772) with poly (A) tail, respectively. Prediction of stem loop structure revealed that, PvFer-IRE can be folded into a typical secondary stem loop structure, having 5-CAGUGA-3' loop, proximal stem of five paired bases followed by a bulged cysteine, and six nucleotide bottom stem, indicating that expression of PvFer is regulated by iron at the translational level. ORF was found to encode 175 amino acid protein with calculated molecular mass of 19.97 kDa and isoelectric point of 4.97. Examination for signal peptide and phylogenetic analysis confirmed that PvFer belonged to cytosolic ferritins of molluscs. Conserved domain analysis showed that PvFer contained both ferroxidase diiron center and ferrihydrite nucleation center, analogous to ferritin M subunit of bony fishes and amphibians. However, amino acid sequence and glycosylation site showed more homology to vertebrate ferritin H subunits. Predicted 3D models of PvFer resembled the typical spatial features of ferritin proteins. The study forms the first comprehensive identification of a ferritin subunit gene in a true/common mussel (Order: Mytilida). Further, the detailed molecular phylogeny conducted through the present study revealed certain thought provoking insights on ferritin genes of the phylum Mollusca.
Assuntos
Ferritinas/genética , Ferritinas/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Perna (Organismo)/genética , Perna (Organismo)/imunologia , Animais , Sequência de Bases , DNA Complementar/análise , Ferritinas/química , Filogenia , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Sediment microbial fuel cells (SMFCs) are bio-electrochemical devices generating electricity from redox gradients occurring across the sediment-water interface. Sediment microbial carbon-capture cell (SMCC), a modified SMFC, uses algae grown in the overlying water of sediment and is considered as a promising system for power generation along with algal cultivation. In this study, the performance of SMCC and SMFC was evaluated in terms of power generation, dissolved oxygen variations, sediment organic matter removal and algal growth. SMCC gave a maximum power density of 22.19 mW/m2, which was 3.65 times higher than the SMFC operated under similar conditions. Sediment organic matter removal efficiencies of 77.6 ± 2.1% and 61.0 ± 1.3% were obtained in SMCC and SMFC, respectively. With presence of algae at the cathode, a maximum chemical oxygen demand and total nitrogen removal efficiencies of 63.3 ± 2.3% (8th day) and 81.6 ± 1.2% (10th day), respectively, were observed. The system appears to be favorable from a resources utilization perspective as it does not depend on external aeration or membranes and utilizes algae and organic matter present in sediment for power generation. Thus, SMCC has proven its applicability for installation in an existing oxidation pond for sediment remediation, algae growth, carbon conversion and power generation, simultaneously.
Assuntos
Fontes de Energia Bioelétrica , Sedimentos Geológicos/química , Microalgas/fisiologia , Água/química , Análise da Demanda Biológica de Oxigênio , Eletricidade , Eletrodos , Nitrogênio , Oxirredução , OxigênioRESUMO
Lately, the bio electrochemical systems are emerging as an efficient wastewater treatment and energy conversion technology. However, their scaling-up is considerably restrained by slow-rate of cathodic oxygen reduction reaction (ORR) or otherwise by the high cost associated with the available efficient ORR catalysts. In this investigation, a cost-effective and eco-friendly approach for synthesizing Ni based ORR catalyst utilizing biosorption property of microalgae is accomplished. The synthesised Ni adsorbed algal biochar (NAB) served as an efficient cathode catalyst for enhancing ORR in a microbial carbon-capture cell (MCC). On increasing the initial concentration of Ni2+ in the aqueous medium from 100 mgL-1 to 500 mgL-1, the biosorption capacity was found to increase from 3 mgg-1 to 32 mgg-1 of algae cell. The MCC operated with NAB based cathode catalyst loading of 2 mgcm-2 exhibited 3.5 times higher power density (4.69 Wm-3) as compared to the one with commercial activated carbon. A significant organic matter removal (82 %) in the anodic chamber with simultaneous algal biomass productivity in the cathodic chamber was attained by MCC with cathode loaded with 2 mgcm-2 of NAB. Hence, this easily synthesised low-cost catalyst, out of waste stream, proved its ability to improve the performance of MCC.
Assuntos
Carvão Vegetal , Níquel , Oxirredução , Oxigênio , Níquel/química , Carvão Vegetal/química , Catálise , Oxigênio/química , Oxigênio/metabolismo , Adsorção , Eletrodos , Microalgas/metabolismo , Biomassa , Fontes de Energia BioelétricaRESUMO
Native lipids in cell-membrane support crucial functions like intercell communication via their ability to deform into curved membrane structures. Cell membrane mimicking Giant unilamellar vesicles (GUV) is imperative in understanding native lipid's role in membrane transformation however remains challenging to assemble. We construct two giant vesicle models mimicking bacterial inner-membrane (IM) and outer-membrane (OM) under physiological conditions using single-step gel-assisted lipid swelling. IM vesicles composed of native bacterial lipids undergo small-scale membrane remodeling into bud and short-nanotube structures. In contrast, OM vesicles asymmetrically assembled from Lipopolysaccharide (LPS) and bacterial lipids underwent global membrane deformation under controlled osmotic stress. Remarkably, highly-curved structures mimicking cell-membrane architectures, including daughter vesicle networks interconnected by necks and nano-tubes ranging from micro to nanoscale, are generated in OM vesicles at osmotic stress comparable to that applied in IM vesicles. Further, we provide a quantitative description of the membrane structures by experimentally determining membrane elastic parameters, i.e., neck curvature and bending rigidity. We can conclude that a larger spontaneous curvature estimated from the neck curvature and softer membranes in OM vesicles is responsible for large-scale deformation compared to IM vesicles. Our findings will help comprehend the shape dynamics of complex native bacterial lipid membranes.
Assuntos
Nanotubos , Lipossomas Unilamelares , Membrana Celular , LipídeosRESUMO
Formation of methane in the anodic chamber of a microbial fuel cell (MFC) indicates an energy inefficiency in electricity generation as the energy required for electrogenesis gets redirected to methanogenesis. The hypothesis of this research is that inhibition of methanogenesis in the mixed anaerobic anodic inoculum is associated with an enhanced activity of the electrogenic bacterial consortia. Hence, the primary objective of this investigation is to evaluate the ability of chloroform to inhibit the methanogenesis at different dosing to enhance the activity of electrogenic consortia in MFC. A higher methane inhibition and hence an enhanced performance of MFC was achieved when mixed anaerobic sludge, collected from septic tank, was used as inoculum after pre-treatment with 0.25% (v/v) chloroform dosing (MFC-0.25CF). The MFC-0.25CF attained a maximum power density of 8.51 W/m3, which was more than twice as that of MFC inoculated with untreated sludge. Also, a clear correlation between the chloroform dosing, methane inhibition, wastewater treatment, and power generation was established, which demonstrated the effectiveness of the technique in enhancing power generation in MFC along with adequate biodegradation of organic matter present in wastewater at an optimum chloroform dosing of 0.25% (v/v) to inhibit methanogenesis.
Assuntos
Fontes de Energia Bioelétrica/microbiologia , Clorofórmio/química , Metano/metabolismo , Esgotos/microbiologia , Anaerobiose , EletrodosRESUMO
The fish market wastewater, which is rich in ammonium concentration, was investigated to explore its ability of in situ suppression of methanogenesis in the anodic chamber of microbial fuel cell (MFC) while treating it and to ensure non-reoccurrence of methanogenic consortia in the anodic chamber during its long-term operations. A lower specific methanogenic activity (0.097g chemical oxygen demand (COD)CH4/g volatile suspended solids (VSS). day) with a higher power density (3.81 ± 0.19 W/m3) was exhibited by the MFC operated with raw fish market wastewater as compared to the MFC fed with synthetic wastewater (0.219g CODCH4/g VSS. day and 1.75 ± 0.09 W/m3, respectively). The enhanced electrochemical activity of anodic biofilm of MFC fed with raw fish market wastewater than the MFC fed with synthetic wastewater further advocated the enhanced electrogenic activity and suppression of methanogenesis, because of the presence of higher ammonium content in the feed. This, in response, reduced the internal resistance (55 Ω), enhanced the coulombic efficiency (21.9 ± 0.3%) and normalized the energy recovery (0.27 kWh/m3) from the MFC fed with fish market wastewater than the MFC fed with synthetic wastewater (92 Ω, 15.7 ± 0.3% and 0.13 kWh/m3, respectively). Thus, while treating the fish market wastewater in the anodic chamber of MFC, any costly and repetitive treatment procedures for anodic microorganisms are not required for suppression of methanogens to ensure higher activity of electrogenic bacteria for higher electricity harvesting.