Microbes, cables, and an electrical touch

In nature, highly efficient and diverse consortia of microbes cycle carbon and other elements while generating energy for growth. Driving these reactions are organisms with the ability to extract electrons from the chemical substrates and transfer them to insoluble and soluble electron acceptors. One bacterial group in particular, Geobacter spp., can couple their respiratory metabolism to the reduction of insoluble minerals, such as iron and manganese oxides, and soluble toxic metals such as uranium. Key to these activities is the ability of the cells to transfer respiratory electrons extracellularly using an electroactive cell envelope containing abundant metalloproteins, including c-cytochromes, and conductive protein appendages or pili (known as nanowires). Thus, in addition to been ecological drivers of the cycling of carbon and metals in nature, these organisms show promise for the bioremediation of environments impacted with toxic metals. The electrical activity of Geobacter can also be mimicked in electrochemical reactors equipped with an electrode poised at a metabolically oxidizing potential, so that the electrode functions as an unlimited sink of electrons to drive the oxidation of electron donors and support cell growth. Electrochemical reactors are promising for the treatments of agricultural, industrial, and human wastes, and the electroactivity of these microbes can be used to develop materials and devices for bioenergy and bioremediation applications. [Int Microbiol 18(3):151-157 (2015)]Keywords: Geobacter · c-cytochromes · electrochemical reactors · microbial fuel cells · nanowires · type IV pil

Electrochemical characterization of Geobacter lovleyi identifies limitations of microbial fuel cell performance in constructed wetlands

Power generation in microbial fuel cells implemented in constructed wetlands (CW-MFCs) is low despite the enrichment of anode electricigens most closely related to Geobacter lovleyi. Using the model representative G. lovleyi strain SZ, we show that acetate, but not formate or lactate, can be oxidized efficiently but growth is limited by the high sensitivity of the bacterium to oxygen. Acetate and highly reducing conditions also supported the growth of anode biofilms but only at optimal anode potentials (450 mV vs. standard hydrogen electrode). Still, electrode coverage was poor and current densities, low, consistent with the lack of key c-type cytochromes. The results suggest that the low oxygen tolerance of G. lovleyi and inability to efficiently colonize and form electroactive biofilms on the electrodes while oxidizing the range of electron donors available in constructed wetlands limits MFC performance. The implications of these findings for the optimization of CW-MFCs are discussed. [Int Microbiol 20(2):55-64 (2017)]Keywords: microbial fuel cells; bioelectrochemical systems; constructed wetlands; extracellular electron transfer; electricigen

Electrochemical characterization of Geobacter lovleyi identifies limitations of microbial fuel cell performance in constructed wetlands

Microbial Fuel Cells implemented in Constructed Wetlands (CW-MFCs) show limited performance. Geobacter Lovleyi has been demonstrated to be one of the predominant bacterial species in active CW-MFCs. The aim of this study was to characterize the growth of G.Lovleyi so as to identify if it could be a source for the observed CW-MFCs low performances. To this aim, G. Lovelyi was grown under three different electron donors (acetate, lactate and formate) and two electron acceptors (fumarate and Fe(III) citrate). G. Lovleyi growing and electron transfer characteristics was also studied by inoculating it in double chambered MECs (anodes poised at 31, 450 and 771 mV). Results showed that its growth was supported by acetate, with doubling times of 4.4±0.1 and 8±0.1 hours for fumarate and Fe(III) citrate as electron acceptors, respectively. G. Lovleyi was also demonstrated to be highly intolerant to oxygen, requiring cysteine as a reducing agent. In contrast, formate and lactate did not support cell growth even in the presence of cysteine. Maximum currents achieved were that of 0.08 mA and 0.26 mA for the MECs operated at 450 mV and 771 mV, respectively. However, no current was observed at 31mV. Confocal laser scanning microscopy (CLSM) analysis showed poor electrode coverage, indicating that G. Lovleyi did not attach to the electrode effectively. According to these results, low performances of CW-MFCs could by at least partially explained by the inability of G. lovleyi to oxidize the wide range of metabolites present in CW, to tolerate even trace oxygen concentrations or to efficiently attach to electrodes surface.Peer ReviewedPostprint (published version

Genome-wide analysis of the RpoN regulon in Geobacter sulfurreducens

Background The role of the RNA polymerase sigma factor RpoN in regulation of gene expression in Geobacter sulfurreducens was investigated to better understand transcriptional regulatory networks as part of an effort to develop regulatory modules for genome-scale in silico models, which can predict the physiological responses of Geobacter species during groundwater bioremediation or electricity production. Results An rpoN deletion mutant could not be obtained under all conditions tested. In order to investigate the regulon of the G. sulfurreducens RpoN, an RpoN over-expression strain was made in which an extra copy of the rpoN gene was under the control of a taclac promoter. Combining both the microarray transcriptome analysis and the computational prediction revealed that the G. sulfurreducens RpoN controls genes involved in a wide range of cellular functions. Most importantly, RpoN controls the expression of the dcuB gene encoding the fumarate/succinate exchanger, which is essential for cell growth with fumarate as the terminal electron acceptor in G. sulfurreducens. RpoN also controls genes, which encode enzymes for both pathways of ammonia assimilation that is predicted to be essential under all growth conditions in G. sulfurreducens. Other genes that were identified as part of the RpoN regulon using either the computational prediction or the microarray transcriptome analysis included genes involved in flagella biosynthesis, pili biosynthesis and genes involved in central metabolism enzymes and cytochromes involved in extracellular electron transfer to Fe(III), which are known to be important for growth in subsurface environment or electricity production in microbial fuel cells. The consensus sequence for the predicted RpoN-regulated promoter elements is TTGGCACGGTTTTTGCT. Conclusion The G. sulfurreducens RpoN is an essential sigma factor and a global regulator involved in a complex transcriptional network controlling a variety of cellular processes

Br J Pharmacol

Background and Purpose : The enzyme α/β-hydrolase domain containing 6 (ABHD6), a new member of the endocannabinoid system, is a promising therapeutic target against neuronal-related diseases. However, how ABHD6 activity is regulated is not known. ABHD6 coexists in protein complexes with the brain-specific carnitine palmitoyltransferase 1C (CPT1C). CPT1C is involved in neuro-metabolic functions, depending on brain malonyl–CoA levels. Our aim was to study CPT1C–ABHD6 interaction and determine whether CPT1C is a key regulator of ABHD6 activity depending on nutritional status. Experimental Approach : Co-immunoprecipitation and FRET assays were used to explore ABHD6 interaction with CPT1C or modified malonyl–CoA-insensitive or C-terminal truncated CPT1C forms. Cannabinoid CB1 receptor-mediated signalling was investigated by determining cAMP levels. A novel highly sensitive fluorescent method was optimized to measure ABHD6 activity in non-neuronal and neuronal cells and in brain tissues from wild-type (WT) and CPT1C–KO mice. Key Results : CPT1C interacted with ABHD6 and negatively regulated its hydrolase activity, thereby regulating 2-AG downstream signalling. Accordingly, brain tissues of CPT1C–KO mice showed increased ABHD6 activity. CPT1C malonyl–CoA sensing was key to the regulatory role on ABHD6 activity and CB1 receptor signalling. Fasting, which attenuates brain malonyl–CoA, significantly increased ABHD6 activity in hypothalamus from WT, but not CPT1C–KO, mice. Conclusions and Implications : Our finding that negative regulation of ABHD6 activity, particularly in the hypothalamus, is sensitive to nutritional status throws new light on the characterization and the importance of the proteins involved as potential targets against diseases affecting the CNS

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Chitin degradation by the facultatively aerobic cellulolytic bacterium Cellulomonas uda

Microbes, traditionally characterized by their capacity to decompose cellulose anaerobically, were examined for the ability to degrade the chemically-related polymer, chitin. Our results suggest that the ability to degrade and utilize chitin as a source of nitrogen might be widespread among cellulolytic bacteria. We suggest that, in natural environments, the ability to degrade chitin may confer an enormous ecological advantage on cellulolytic microorganisms as chitin may serve as a source of nitrogen, which is often limiting in environments where cellulose accumulates. The soil isolate, Cellulomonas uda ATCC 21399, was selected for further studies inasmuch as cultures of this strain rapidly degraded cellulose and chitin, aerobically and anaerobically, and active cellulases and chitinases were present in culture supernatant fluids. Our studies indicate that the cellulase and chitinase systems of C. uda are distinct based on differences in their protein composition and the regulation of their production by growth substrates. C. uda efficiently degraded chitin with a relatively simple chitinase system composed of a major enzyme component, an endochitinase designated ChiA, and one or more exochitinases. ChiA was purified from culture supernatant fluids and characterized. It was found to be a glycoprotein with a Mr of 70kDa, pI of 8.0 and an endochitinase mode of action, ChiA degraded chitinous substrates with different degrees of crystallinity and strongly bound to both chitinous and cellulosic substrates. Optimum temperature for activity against colloidal chitin was 60°C, and optimum pH was 7.0. Preliminary information on the nucleotide sequence of chiA, the gene coding for ChiA of C. uda, suggested that ChiA might be closely related to chitinases from Streptomyces species, such as ChiC from Streptomyces lividans. The influence of nitrogen availability on growth of C. uda was studied, C. uda cells grew as a biofilm in response to conditions of nitrogen limitation. Addition of sources of nitrogen, including products of chitin degradation, resulted in biofilm detachment. The biofilms attached to biotic surfaces such as cellulose and chitin and appeared to be involved in their degradation. Chitinase activity was detected in supernatant fluids from cultures where cells were growing as a biofilm, suggesting that nitrogen availability might also regulate the production of chitinases. Our results suggest that bacterial biofilms play an important role in the degradation of polymers in nature

Virulence and the Environment: a Novel Role for Vibrio cholerae Toxin-Coregulated Pili in Biofilm Formation on Chitin

The toxin-coregulated pilus (TCP) of Vibrio cholerae is required for intestinal colonization and cholera toxin acquisition. Here we report that TCP mediates bacterial interactions required for biofilm differentiation on chitinaceous surfaces. We also show that undifferentiated TCP(−) biofilms have reduced ecological fitness and, thus, that chitin colonization may represent an ecological setting outside the host in which selection for a host colonization factor may take place