Biodegradation of diesel fuel hydrocarbons by mangrove fungi from Red Sea Coast of Saudi Arabia

Mangrove sediments were collected from major mangrove stands on the Red Sea Coast of Saudi Arabia. Forty five isolates belonging to 12 genera were purified and five isolates as well as their consortium were found to be able to grow in association with petroleum oil as sole carbon source under in vitro conditions. The isolated strains were identified based on internal transcribed spacer (ITS) rDNA sequence analysis. The fungal strains with the greatest potentiality to degrade diesel oil, without developing antagonistic activity, were identified as Alternaria alternata, Aspergillus terreus, Cladosporium sphaerospermum, Eupenicillium hirayamae and Paecilomyces variotii. As compared to the controls, these fungi accumulated significantly higher biomass, produced extracellular enzymes and liberated larger volumes of CO₂. These observations with GC–MS data confirm that these isolates displayed rapid diesel oil bioremoval and when used together as a consortium, there was no antagonistic activity.     

Enhanced biodegradation of phenanthrene using different inoculum types in a microbial fuel cell

Environmental pollution by petroleum hydrocarbons from contaminated groundwater and soils is a serious threat to human health. Microbial fuel cells (MFCs) could be employed in the treatment of these recalcitrant pollutants with concomitant bioelectricity generation. In this study, the use of MFCs in biodegradation of phenanthrene, a model hydrocarbon, was investigated with respect to its biodegradation rate, biodegradation efficiency, and power production using a range of inocula (Shewanella oneidensis MR1 14063, Pseudomonas aeruginosa NCTC 10662, mixed cultures, and combinations thereof). All the inocula showed high potentials for phenanthrene degradation with a minimum degradation efficiency of 97%. The best overall performing inoculum was anaerobically digested sludge supplemented with P. aeruginosa NCTC 10662, having a degradation rate, maximum power density and chemical oxygen demand removal efficiency of 27.30 μM/d, 1.25 mW/m² and 65.6%, respectively. Adsorption of phenanthrene on the carbon anode was also investigated; it conformed to a Type II adsorption isotherm and could be modelled using a modified Brunauer, Emmett and Teller model with a maximum monolayer capacity of 0.088 mg/cm². This work highlights the possibility of using MFCs to achieve high degradation rates of phenanthrene through co‐metabolism and could potentially be used as a replacement of permeable reactive barriers for remediation of hydrocarbon‐contaminated groundwater.     

Change in microbial communities in acetate- and glucose-fed microbial fuel cells in the presence of light

Power densities produced by microbial fuel cells (MFCs) in natural systems are changed by exposure to light through the enrichment of photosynthetic microorganisms. When MFCs with brush anodes were exposed to light (4000 lx), power densities increased by 8–10% for glucose-fed reactors, and 34% for acetate-fed reactors. Denaturing gradient gel electrophoresis (DGGE) profiles based on the 16S rRNA gene showed that exposure to high light levels changed the microbial communities on the anodes. Based on 16S rRNA gene clone libraries of light-exposed systems the anode communities using glucose were also significantly different than those fed acetate. Dominant bacteria that are known exoelectrogens were identified in the anode biofilm, including a purple nonsulfur (PNS) photosynthetic bacterium, Rhodopseudomonas palustris, and a dissimilatory iron-reducing bacterium, Geobacter sulfurreducens. Pure culture tests confirmed that PNS photosynthetic bacteria increased power production when exposed to high light intensities (4000 lx). These results demonstrate that power production and community composition are affected by light conditions as well as electron donors in single-chamber air-cathode MFCs.     

Degradation of pentachlorophenol with the presence of fermentable and non-fermentable co-substrates in a microbial fuel cell

Pentachlorophenol (PCP) was more rapidly degraded in acetate and glucose-fed microbial fuel cells (MFCs) than in open circuit controls, with removal rates of 0.12 ± 0.01 mg/Lh (14.8 ± 1.0 mg/g-VSS-h) in acetate-fed, and 0.08 ± 0.01 mg/L h (6.9 ± 0.8 mg/g-VSS-h) in glucose-fed MFCs, at an initial PCP concentration of 15 mg/L. A PCP of 15 mg/L had no effect on power generation from acetate but power production was decreased with glucose. Coulombic balances indicate the predominant product was electricity (16.1 ± 0.3%) in PCP-acetate MFCs, and lactate (19.8 ± 3.3%) in PCP-glucose MFCs. Current generation accelerated the removal of PCP and co-substrates, as well as the degradation products in both PCP-acetate and PCP-glucose reactors. While 2,3,4,5-tetrachlorophenol was present in both reactors, tetrachlorohydroquinone was only found in PCP-acetate MFCs. These results demonstrate PCP degradation and power production were affected by current generation and the type of electron donor provided.     

Utilization of proteinaceous materials for power generation in a mediatorless microbial fuel cell by a new electrogenic bacteria Lysinibacillus sphaericus VA5

In this study, a bacterial strain, Lysinibacillus sphaericus which is relatively new in the vast list of biocatalysts known to produce electricity has been tested for its potential in power production. It is cited from the literature that the organism is deficient in some sugar or polysaccharide processing enzymes and thus is tested for its ability to utilize substrates mainly rich in protein components like beef extract and with successive production of electricity. The particular species has been found to generate a maximum power density of 85 mW/m² and current density of ≈270 mA/m² using graphite felt as electrode. The maximum Open Circuit Voltage and current has been noted as 0.7 Vand 0.8 mA during these operational cycles. Cyclic voltammetry studies indicate the presence of some electroactive compounds which can facilitate electron transfer from bacteria to electrode. The number of electrogens able to generate electricity in mediator free conditions are few, and the study introduces more divergence to that population. Substrate specificity and electricity generation efficacy of the strain in treating wastewater, specially rich in protein content has been reported in the study. As the species has been found to be efficient in utilizing proteinaceous material, the technique can be useful to treat specific type of wastewaters like wastewater from slaughterhouses or from meat packaging industry. Treating them in a more economical way which generates electricity as a outcome must be preferred over the conventional aerobic treatments. Emphasizing on substrate specificity, the study introduces this novel Lysinibacillus strain as a potent biocatalyst and its sustainable role in MFC application for bioenergy generation.     

Enhancement of coulombic efficiency and salt tolerance in microbial fuel cells by graphite/alginate granules immobilization of Shewanella oneidensis MR-1

Coulombic efficiency and stability of electricity output are crucial for practical applications of microbial fuel cells (MFCs). In this study, a cell immobilization method for electrogenic microorganism in MFCs using graphite/alginate granules is developed. The MFC with immobilized cell granules delivered a much more stable electricity output than that with suspension cells, and resulted in a ～0.8 to 1.7 times improvement on coulombic efficiency compared to the suspension mode. Impressively, with the conductive graphite/alginate/cells granules, the internal resistance of the MFC decreased dramatically. Moreover, the cell immobilized MFC showed a much higher tolerance to the shock of high salt concentration than the MFC with suspension cells. The results substantiated that immobilization of electrogenic microorganism for MFCs could be achieved by the method developed here, and it is promising for practical application in energy harvesting from wastewater by MFCs.     

Biodegradation of fuel oxygenates and their effect on the expression of a newly identified cytochrome P450 gene in Achromobacter xylosoxidans MCM2/2/1

Achromobacter xylosoxidans MCM2/2/1 was enriched and isolated from gasoline-contaminated soil and was found to degrade ethyl tert-butyl ether (ETBE) and methyl tert-butyl ether (MTBE) by 41.48% and 34.15%, respectively, in 6 days. Furthermore, the effect of MTBE and TBA on the expression of cytochrome P450 (CYP) of A. xylosoxidans MCM2/2/1 was examined. The presence of the CYP gene in this organism was first confirmed by amplification of a putative 350 bp CYP gene fragment followed by identification of the entire gene by genome walking and DNA-sequencing. The identified CYP gene of A. xylosoxidans MCM2/2/1 shares a high similarity of about 88% with the thcB gene of A. xylosoxidans A8. Gene expression studies have shown that the CYP gene is expressed in A. xylosoxidans MCM2/2/1; however, the expression of this gene was altered at different concentrations of MTBE.     

Biodegradation of carbazole in oil/water biphasic system by a newly isolated bacterium Klebsiella sp. LSSE-H2

A novel Klebsiella sp. strain LSSE-H2 (CGMCC No. 1624) was isolated from dye-contaminated soil based on its ability to metabolize carbazole as a sole source of carbon and nitrogen. This strain efficiently degraded carbazole from either aqueous and biphasic aqueous–organic media, displaying a high denitrogenation activity and a high level of solvent tolerance. LSSE-H2 could completely degrade 12 mmol/L carbazole after 56 h of cultivation. The co-culture of LSSE-H2 and Pseudomonas delafieldii R-8 strains can degrade approximately 92% of carbazole (10 mmol/L) and 94% of dibenzothiophene (3 mmol/L) from model diesel in 12 h.     

The direct electrocatalysis of phenazine-1-carboxylic acid excreted by Pseudomonas alcaliphila under alkaline condition in microbial fuel cells

In this paper, we reported a kind of exoelectrogens, Pseudomonas alcaliphila (P. alcaliphila) strain MBR, which could excrete phenazine-1-carboxylic acid (PCA) to transfer electron under alkaline condition in microbial fuel cells (MFCs). The electrochemical activity of strain MBR and the extracellular electron transfer mechanism in MFCs were evaluated by cyclic voltammetry (CV) and electricity generation curve measurement. The results indicated a soluble mediator was the key factor for extracellular electron transfer of strain MBR under alkaline condition. The soluble mediator was PCA detected by gas chromatography-mass (GC-MS) analyses.     

Understanding the role of Fe(III)/Fe(II) couple in mediating reductive transformation of 2-nitrophenol in microbial fuel cells

The Fe(III)/Fe(II) couple can play a significant role in the abiotic reduction of 2-nitrophenol (2-NP) at the cathode chamber of a microbial fuel cell (MFC). Experimental results demonstrate that Fe(II) addition to the cathode chamber contributes to a significant increase in the reaction rate of 2-NP removal and the power performance of MFC. Observed pseudo first-order rate constants and power densities are heavily dependent on the identity of the Fe(II)-complexing ligands. The Fe(II) complex coordinated with citrate results in the highest rate constant up to 0.12 h⁻¹ as compared to other organically complexed iron species including Fe(II)-EDTA, Fe(II)-acetate and Fe(II)-oxalate, and iron species uncomplexed with any organic ligands. In addition, the presence of Fe(II)-citrate species leads to a maximum volumetric power density of 1.0 W m⁻³, which is the highest value among those obtained with other iron species for the similar MFC system.     

Metabolism of kaurenoids by Gibberella fujikuroi in the presence of the plant growth retardant,N,N,N-trimethyl-1-methyl-(2′,6′,6′-trimethylcyclohex-2′-en-1′-yl)prop-2-enylammonium iodide

The plant growth retardant, N,N,N-trimethyl-1-methyl-(2′,6′,6′-trimethylcyclohex-2′-en-1′-yl)prop-2-enylammonium iodide, is shown to block gibberellin biosynthesis in Gibberella fujikuroi between mevalonate and ent-kaur-16-ene, probably by inhibiting ent-kaur-16-ene synthetase A-activity. In the presence of the plant growth retardant, cultures of the fungus incorporate (26.5%) added ent-[¹⁴C]-kaur-16-ene into gibberellin A₃. Under the same conditions kaur-16-ene, 13β-kaur-16-ene, and ent-kaur-15-ene are not metabolised to gibberellin analogues.     

Biodegradation of methyl parathion and <Emphasis Type="Italic">p</Emphasis>-nitrophenol: evidence for the presence of a <Emphasis Type="Italic">p</Emphasis>-nitrophenol 2-hydroxylase in a Gram-negative <Emphasis Type="Italic">Serratia</Emphasis> sp. strain DS001

A soil bacterium capable of utilizing methyl parathion as sole carbon and energy source was isolated by selective enrichment on minimal medium containing methyl parathion. The strain was identified as belonging to the genus Serratia based on a phylogram constructed using the complete sequence of the 16S rRNA. Serratia sp. strain DS001 utilized methyl parathion, p-nitrophenol, 4-nitrocatechol, and 1,2,4-benzenetriol as sole carbon and energy sources but could not grow using hydroquinone as a source of carbon. p-Nitrophenol and dimethylthiophosphoric acid were found to be the major degradation products of methyl parathion. Growth on p-nitrophenol led to release of stoichiometric amounts of nitrite and to the formation of 4-nitrocatechol and benzenetriol. When these catabolic intermediates of p-nitrophenol were added to resting cells of Serratia sp. strain DS001 oxygen consumption was detected whereas no oxygen consumption was apparent when hydroquinone was added to the resting cells suggesting that it is not part of the p-nitrophenol degradation pathway. Key enzymes involved in degradation of methyl parathion and in conversion of p-nitrophenol to 4-nitrocatechol, namely parathion hydrolase and p-nitrophenol hydroxylase component “A” were detected in the proteomes of the methyl parathion and p-nitrophenol grown cultures, respectively. These studies report for the first time the existence of a p-nitrophenol hydroxylase component “A”, typically found in Gram-positive bacteria, in a Gram-negative strain of the genus Serratia. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Characterization of 1-(2-[18F] fluoro-3-pyridyl)-4-(2-isopropyl-1-oxo- isoindoline-5-yl)-5-methyl-1H-1,2,3-triazole, a PET ligand for imaging the metabotropic glutamate receptor type 1 in rat and monkey brains

Neurovascular degeneration contributes to the pathogenesis of Alzheimer's disease (AD). Because erythropoietin (EPO) promotes endothelial regeneration, we investigated the therapeutic effects of EPO in animal models of AD. In aged Tg2576 mice, EPO receptors (EPORs) were expressed in the cortex and hippocampus. Tg2576 mice were treated with daily injection of EPO (5000 IU/kg/day) for 5 days. At 14 days, EPO improved contextual memory as measured by fear-conditioning test. EPO enhanced endothelial proliferation and the level of synaptophysin expression in the brain. EPO also increased capillary density, and decreased the level of the receptor for advanced glycation endproducts (RAGE) in the brain, while decreasing in the amount of amyloid plaque and amyloid-β (Aβ). In cultured human endothelial cells, EPO enhanced angiogenesis and suppressed the expression of the RAGE. These results show that EPO improves memory and ameliorates endothelial degeneration induced by Aβ in AD models. This pre-clinical evidence suggests that EPO may be useful for the treatment of AD.     

Conversion of the 2 Cl−/1 H+ antiporter ClC‐5 in a NO3−/H+ antiporter by a single point mutation

Several members of the CLC family are secondary active anion/proton exchangers, and not passive chloride channels. Among the exchangers, the endosomal ClC-5 protein that is mutated in Dent''s disease shows an extreme outward rectification that precludes a precise determination of its transport stoichiometry from measurements of the reversal potential. We developed a novel imaging method to determine the absolute proton flux in Xenopus oocytes from the extracellular proton gradient. We determined a transport stoichiometry of 2 Cl⁻/1 H⁺. Nitrate uncoupled proton transport but mutating the highly conserved serine 168 to proline, as found in the plant NO₃⁻/H⁺ antiporter atClCa, led to coupled NO₃⁻/H⁺ exchange. Among several amino acids tested at position 168, S168P was unique in mediating highly coupled NO₃⁻/H⁺ exchange. We further found that ClC-5 is strongly stimulated by intracellular protons in an allosteric manner with an apparent pK of ∼7.2. A 2:1 stoichiometry appears to be a general property of CLC anion/proton exchangers. Serine 168 has an important function in determining anionic specificity of the exchange mechanism.     

Insights into the binding specificity of wild type and mutated wheat germ agglutinin towards Neu5Acα(2‐3)Gal: a study by in silico mutations and molecular dynamics simulations

Wheat germ agglutinin (WGA) is a plant lectin, which specifically recognizes the sugars NeuNAc and GlcNAc. Mutated WGA with enhanced binding specificity can be used as biomarkers for cancer. In silico mutations are performed at the active site of WGA to enhance the binding specificity towards sialylglycans, and molecular dynamics simulations of 20 ns are carried out for wild type and mutated WGAs (WGA1, WGA2, and WGA3) in complex with sialylgalactose to examine the change in binding specificity. MD simulations reveal the change in binding specificity of wild type and mutated WGAs towards sialylgalactose and bound conformational flexibility of sialylgalactose. The mutated polar amino acid residues Asn114 (S114N), Lys118 (G118K), and Arg118 (G118R) make direct and water mediated hydrogen bonds and hydrophobic interactions with sialylgalactose. An analysis of possible hydrogen bonds, hydrophobic interactions, total pair wise interaction energy between active site residues and sialylgalactose and MM‐PBSA free energy calculation reveals the plausible binding modes and the role of water in stabilizing different binding modes. An interesting observation is that the binding specificity of mutated WGAs (cyborg lectin) towards sialylgalactose is found to be higher in double point mutation (WGA3). One of the substituted residues Arg118 plays a crucial role in sugar binding. Based on the interactions and energy calculations, it is concluded that the order of binding specificity of WGAs towards sialylgalactose is WGA3 > WGA1 > WGA2 > WGA. On comparing with the wild type, double point mutated WGA (WGA3) exhibits increased specificity towards sialylgalactose, and thus, it can be effectively used in targeted drug delivery and as biological cell marker in cancer therapeutics. Copyright © 2014 John Wiley & Sons, Ltd.