4株高效产电菌的电化学活性及其生物学特征

采用循环伏安法对从湖南省吉首大田湾污水处理厂曝气池活性污泥中富集和筛选的37株产电菌的电化学活性进行考察。结果发现,4株菌株（F012、F015、F021、F026）的电化学活性较为显著,其中F026的电化学活性最好。对4株产电菌的系统发育分析表明,菌株F012属于Dyella属,与Dyella marensis CS5-B2^T系统发育关系最为密切（相似性为97.22%）;菌株F015属于Paludibacterium属,与该属的Paludibacterium yongneupense 5YN8-15^T系统发育关系最为密切（相似性为97.70%）;菌株F021和F026都属于Pseudomonas属,分别与该属的Pseudomonas simiae OLi^T和Pseudomonas otitidis MCC10330^T系统发育关系最为密切（相似性分别为99.60%和98.62%）。生物学特性研究表明,电化学活性最好的产电菌F026的生长温度范围为20~40 ℃,最适宜生长温度为30~35 ℃;生长pH范围为5~9,最适pH生长范围为8~9,适合作为微生物燃料电池的高效产电菌。     

Continuous power generation and microbial community structure of the anode biofilms in a three-stage microbial fuel cell system

A mediator-less three-stage two-chamber microbial fuel cell (MFC) system was developed and operated continuously for more than 1.5 years to evaluate continuous power generation while treating artificial wastewater containing glucose (10 mM) concurrently. A stable power density of 28 W/m³ was attained with an anode hydraulic retention time of 4.5 h and phosphate buffer as the cathode electrolyte. An overall dissolved organic carbon removal ratio was about 85%, and coulombic efficiency was about 46% in this MFC system. We also analyzed the microbial community structure of anode biofilms in each MFC. Since the environment in each MFC was different due to passing on the products to the next MFC in series, the microbial community structure was different accordingly. The anode biofilm in the first MFC consisted mainly of bacteria belonging to the Gammaproteobacteria, identified as Aeromonas sp., while the Firmicutes dominated the anode biofilms in the second and third MFCs that were mainly fed with acetate. Cyclic voltammetric results supported the presence of a redox compound(s) associated with the anode biofilm matrix, rather than mobile (dissolved) forms, which could be responsible for the electron transfer to the anode. Scanning electron microscopy revealed that the anode biofilms were comprised of morphologically different cells that were firmly attached on the anode surface and interconnected each other with anchor-like filamentous appendages, which might support the results of cyclic voltammetry. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The Indispensable N-Terminal Half of eIF3j/HCR1 Cooperates with its Structurally Conserved Binding Partner eIF3b/PRT1-RRM and with eIF1A in Stringent AUG Selection

Despite recent progress in our understanding of the numerous functions of individual subunits of eukaryotic translation initiation factor (eIF) 3, little is known on the molecular level. Using NMR spectroscopy, we determined the first solution structure of an interaction between eIF3 subunits. We revealed that a conserved tryptophan residue in the human eIF3j N-terminal acidic motif (NTA) is held in the helix α1 and loop 5 hydrophobic pocket of the human eIF3b RNA recognition motif (RRM). Mutating the corresponding “pocket” residues in its yeast orthologue reduces cellular growth rate, eliminates eIF3j/HCR1 association with eIF3b/PRT1 in vitro and in vivo, affects 40S occupancy of eIF3, and produces a leaky scanning defect indicative of a deregulation of the AUG selection process. Unexpectedly, we found that the N-terminal half of eIF3j/HCR1 containing the NTA is indispensable and sufficient for wild-type growth of yeast cells. Furthermore, we demonstrate that deletion of either j/HCR1 or its N-terminal half only, or mutation of the key tryptophan residues results in the severe leaky scanning phenotype partially suppressible by overexpressed eIF1A, which is thought to stabilize properly formed preinitiation complexes at the correct start codon. These findings indicate that eIF3j/HCR1 remains associated with the scanning preinitiation complexes and does not dissociate from the small ribosomal subunit upon mRNA recruitment, as previously believed. Finally, we provide further support for earlier mapping of the ribosomal binding site for human eIF3j by identifying specific interactions of eIF3j/HCR1 with small ribosomal proteins RPS2 and RPS23 located in the vicinity of the mRNA entry channel. Taken together, we propose that eIF3j/HCR1 closely cooperates with the eIF3b/PRT1 RRM and eIF1A on the ribosome to ensure proper formation of the scanning-arrested conformation required for stringent AUG recognition.     

A copper‐specific microbial fuel cell biosensor based on riboflavin biosynthesis of engineered Escherichia coli

Copper pollution poses a serious threat to the aquatic environment; however, in situ analytical methods for copper monitoring are still scarce. In the current study, Escherichia coli Rosetta was genetically modified to express OprF and ribB with promoter P_t7 and P_cusC, respectively, which could synthesize porin and senses Cu²⁺ to produce riboflavin. The cell membrane permeability of this engineered strain was increased and its riboflavin production (1.45–3.56 μM) was positively correlated to Cu²⁺ (0–0.5 mM). The biosynthetic strain was then employed in microbial fuel cell (MFC) based biosensor. Under optimal operating parameters of pH 7.1 and 37°C, the maximum voltage (248, 295, 333, 352, and 407 mV) of the constructed MFC biosensor showed a linear correlation with Cu²⁺ concentration (0.1, 0.2, 0.3, 0.4, 0.5 mM, respectively; R² = 0.977). The continuous mode testing demonstrated that the MFC biosensor specifically senses Cu²⁺ with calculated detection limit of 28 μM, which conforms to the common Cu²⁺ safety standard (32 μM). The results obtained with the developed biosensor system were consistent with the existing analytical methods such as colorimetry, flame atomic absorption spectrometry, and inductively coupled plasma optical emission spectrometry. In conclusion, this MFC‐based biosensor overcomes the signal conversion and transmission problems of conventional approaches, providing a fast and economic analytical alternative for in situ monitoring of Cu²⁺ in water.     

Microbial fuel cell based biosensor for in situ monitoring of anaerobic digestion process

A wall-jet microbial fuel cell (MFC) was developed for the monitoring of anaerobic digestion (AD). This biofilm based MFC biosensor had a character of being portable, short hydraulic retention time (HRT) for sample flow through and convenient for continuous operation. The MFC was installed in the recirculation loop of an upflow anaerobic fixed-bed (UAFB) reactor in bench-scale where pH of the fermentation broth and biogas flow were monitored in real time. External disturbances to the AD were added on purpose by changing feedstock concentration, as well as process configuration. MFC signals had good correlations with online measurements (i.e. pH, gas flow rate) and offline analysis (i.e. COD) over 6-month operation. These results suggest that the MFC signal can reflect the dynamic variation of AD and can potentially be a valuable tool for monitoring and control of bioprocess.     

Metabolite analysis of Clostridium acetobutylicum: Fermentation in a microbial fuel cell

Microbial fuel cells (MFCs) were used to monitor metabolism changes in Clostridium acetobutylicum fermentations. When MFCs were inoculated with C. acetobutylicum, they generated a unique voltage output pattern where two distinct voltage peaks occurred over a weeklong period. This result was markedly different to previously studied organisms which usually generate one sustained voltage peak. Analysis of the fermentation products indicated that the dual voltage peaks correlated with glucose metabolism. The first voltage peak correlated with acidogenic metabolism (acetate and butyrate production) and the second peak with solventogenic metabolism (acetone and butanol production). This demonstrates that MFCs can be applied as a novel tool to monitor the shift from acid production to solvent production in C. acetobutylicum.     

Functional stability of a hybrid anaerobic digester/microbial fuel cell system treating municipal wastewater

A thermophilic anaerobic digester (AD) was combined with a microbial fuel cell (MFC) to evaluate whether either component had increased stability when operated in combination as a hybrid system, perturbed by the addition of acetic acid. The MFC and the anaerobic digester were able to operate effectively together. The MFC was more susceptible to high acetic acid load than the AD. The hybrid system did not have increased resilience compared to the solitary systems in the conditions tested. However, the low pH had a relatively delayed effect on the MFC compared to the AD, allowing the hybrid system to have a more stable energy output. Also, at very low pH, when operating as a hybrid, the AD component was able to recover pH to normal levels when the MFC component failed. These results demonstrate that there are synergies that can be gained from this hybrid system.     

Effects of biomass weight and light intensity on the performance of photosynthetic microbial fuel cells with Spirulina platensis

Microalgae Spirulina platensis were attached to the anode of a membrane-free and mediator-free microbial fuel cell (MFC) to produce electricity through the consumption of biochemical compounds inside the microalgae. An increase in open circuit voltage (OCV) was observed with decreasing light intensity and optimal biomass area density. The highest OCV observation for the MFC was 0.39 V in the dark with a biomass area density on the anode surface of 1.2 g cm⁻². Additionally, it was observed that the MFC with 0.75 g cm⁻² of biomass area density produced 1.64 mW m⁻² of electrical power in the dark, which is superior to the 0.132 mW m⁻² produced in the light. Which also means the MFC can be applied to generate electrical power under both day and night conditions.     

Change in nutritional status modulates the abundance of critical pre-initiation intermediate complexes during translation initiation in vivo

In eukaryotic translation initiation, eIF2GTP-Met-tRNA(i)(Met) ternary complex (TC) interacts with eIF3-eIF1-eIF5 complex to form the multifactor complex (MFC), while eIF2GDP associates with eIF2B for guanine nucleotide exchange. Gcn2p phosphorylates eIF2 to inhibit eIF2B. Here we evaluate the abundance of eIFs and their pre-initiation intermediate complexes in gcn2 deletion mutant grown under different conditions. We show that ribosomes are three times as abundant as eIF1, eIF2 and eIF5, while eIF3 is half as abundant as the latter three and hence, the limiting component in MFC formation. By quantitative immunoprecipitation, we estimate that approximately 15% of the cellular eIF2 is found in TC during rapid growth in a complex rich medium. Most of the TC is found in MFC, and important, approximately 40% of the total eIF2 is associated with eIF5 but lacks tRNA(i)(Met). When the gcn2Delta mutant grows less rapidly in a defined complete medium, TC abundance increases threefold without altering the abundance of each individual factor. Interestingly, the TC increase is suppressed by eIF5 overexpression and Gcn2p expression. Thus, eIF2B-catalyzed TC formation appears to be fine-tuned by eIF2 phosphorylation and the novel eIF2/eIF5 complex lacking tRNA(i)(Met).