Addition Of Electron Donors Research Articles

Anaerobic Faecalicatena spp. degrade sulfoquinovose via a bifurcated 6-deoxy-6-sulfofructose transketolase/transaldolase pathway to both C2- and C3-sulfonate intermediates

Plant-produced sulfoquinovose (SQ, 6-deoxy-6-sulfoglucose) is one of the most abundant sulfur-containing compounds in nature and its bacterial degradation plays an important role in the biogeochemical sulfur and carbon cycles and in all habitats where SQ is produced and degraded, particularly in gut microbiomes. Here, we report the enrichment and characterization of a strictly anaerobic SQ-degrading bacterial consortium that produces the C2-sulfonate isethionate (ISE) as the major product but also the C3-sulfonate 2,3-dihydroxypropanesulfonate (DHPS), with concomitant production of acetate and hydrogen (H2). In the second step, the ISE was degraded completely to hydrogen sulfide (H2S) when an additional electron donor (external H2) was supplied to the consortium. Through growth experiments, analytical chemistry, genomics, proteomics, and transcriptomics, we found evidence for a combination of the 6-deoxy-6-sulfofructose (SF) transketolase (sulfo-TK) and SF transaldolase (sulfo-TAL) pathways in a SQ-degrading Faecalicatena-phylotype (family Lachnospiraceae) of the consortium, and for the ISE-desulfonating glycyl-radical enzyme pathway, as described for Bilophila wadsworthia, in an Anaerospora-phylotype (Sporomusaceae). Furthermore, using total proteomics, a new gene cluster for a bifurcated SQ pathway was also detected in Faecalicatena sp. DSM22707, which grew with SQ in pure culture, producing mainly ISE, but also 3-sulfolacate (SL) 3-sulfolacaldehyde (SLA), acetate, butyrate, succinate, and formate, but not H2. We then reproduced the growth of the consortium with SQ in a defined co-culture model consisting of Faecalicatena sp. DSM22707 and Bilophila wadsworthia 3.1.6. Our findings provide the first description of an additional sulfoglycolytic, bifurcated SQ pathway. Furthermore, we expand on the knowledge of sulfidogenic SQ degradation by strictly anaerobic co-cultures, comprising SQ-fermenting bacteria and cross-feeding of the sulfonate intermediate to H2S-producing organisms, a process in gut microbiomes that is relevant for human health and disease.

Unveiling the mechanism underlying in−situ enhancement on anammox system by sulfide: Integration of biological and isotope analysis

The in−situ utilization of sulfide to remove the nitrate produced during the anaerobic ammonium oxidation (anammox) process can avoid prolonged sludge acclimatization, facilitating the rapid initiation of coupled nitrogen removal processes. However, the understanding of in−situ enhancement on anammox system by sulfide remains unclear. Herein, sulfide (Na2S) was introduced as an additional electron donor to remove the nitrate derived from the anammox under varying sulfide/nitrogen (S/N, S2−-S/NO3–-N, molar ratio) ratios (0.004–4.375). The underlying mechanisms were elucidated by molecular biology techniques including flow cytometry, quantitative polymerase chain reaction, and 16S rRNA amplicon sequencing, alongside isotope tracer analysis. Results revealed that anammox reactors, when operated with in−situ sulfide addition, exhibited a significant enhancement in total nitrogen removal efficiency (NRE) ranging from 11.5%−41.7% (achieved 96%), with the optimal S/N ratios of 0.01−0.8. Isotope tracer analysis indicated the successful coupling of the anammox, sulfur autotrophic denitrification (SADN), and dissimilatory nitrate reduction to ammonium (DNRA) processes within the system, with their contributions to nitrogen removal being 46%−50%, 24%−30%, and 20%−22%, respectively. Moreover, a notable increase in the abundance of sulfur–oxidizing bacteria (SOB) (20%−40% increase) and DNRA bacteria (10%−20% increase) were observed. Effective collaboration was further supported by the sustained viability of microbial communities. It is speculated that the heightened presence of SOB and DNRA bacteria created a low toxicity environment by converting sulfide to biogenic sulfur, thereby promoting the well−being of anammox bacteria. However, the excessive dosage of sulfide (S/N=1.8) intensified the DNRA process (contribution>35%) and weakened the anammox process, leading to an increase in effluent NH4+-N concentration and a decline in NRE. This study confirms that the in−situ adding an appropriate amount of sulfide favors achieving complete nitrogen removal in anammox system, which provides a novel avenue to resolve the issue of the residual nitrate in anammox process.

Field study on the dynamics of microbial communities following biostimulation at organochlorine-contaminated sites

Chlorinated ethenes (CEs) have become widespread contaminants in soils, sediments, and groundwater due to extensive historical use, improper handling, and disposal. At many sites, biostimulation is an effective remediation strategy for treating CEs, involving the addition of electron donors to support bacteria capable of degrading these compounds. This study aimed to understand how microbial communities adapt to environmental changes induced by biostimulation interventions. We investigated changes in microbial communities on biomass carriers transferred from a non-biostimulated, CE-contaminated environment to a previously biostimulated environment at two sites. In the non-biostimulated wells, a predominance of bacteria with nitrate-reducing or iron-reducing metabolisms was evident. Some of which are known to aerobically degrade CEs or other organic pollutants. Following the transfer of the carriers, these bacteria were outcompeted by anaerobic microorganisms with respiratory and fermentative metabolisms. The newly formed microbial community on the transferred biomass carriers exhibited greater diversity and higher abundance of organohalide-respiring bacteria. This rapid adaptability indicates the potential for these bacteria to flourish in favorable conditions, particularly at Site 2, where five days after carrier transfer, Dehalococcoides abundance increased 14,200-fold, and the biomarker vcrA increased 28,500-fold. PCoA revealed that that greater differences in hydrogeochemical conditions between biostimulated and non-biostimulated wells resulted in accelerated changes in bacterial community composition and diversity. In conclusion, our study demonstrates a rapid proliferation of CE-degrading bacteria after the transfer of biomass carriers into biostimulated wells at two different sites. The transfer stimulated shifts toward organohalide-respiring and other anaerobic bacteria in biostimulated environment and increased microbial diversity.

Recovery of platinum nanoparticles by the anaerobic mixed consortium: Platinum (IV) ion reduction, microbial community dynamics and functional genes identification

Removal and recovery of platinum (Pt) from platinum-contaminated wastestreams is beneficial to tackle the increasing supply risk and prevent adverse environmental impacts. Reduction soluble Pt (IV) to Pt nanoparticles using biological method is a plausible approach. However, current studies mainly focus on pure bacterial cultures to reduce Pt (IV). This study evaluates Pt (IV) removal via bioreduction by anaerobic granular sludge with the addition of exogenous electron donors and elucidates underlying mechanisms. Results show that formate and ethanol exhibited significant effects of Pt (IV) reduction, while acetate, lactate and pyruvate showed limited effects. The reduced products Pt nanoparticles were deposited on bacterial cell surface and periplasm. Moreover, Bacteroidetes, Patescibacteria and Proteobacteria were dominant bacteria during the Pt (IV) reduction process. Pt (IV) reduction was mainly mediated by enzymatic processes. Overall, these findings deepen understanding of Pt (IV) bioreduction by anaerobic mixed consortium.

Characterization of groundwater and cores in the decommissioned acid in-situ leach uranium mining area: Enlightenment for uranium contaminated groundwater remediation

Acid In-situ Leach (AISL) is predominant technique for uranium mining in sandstone-type deposits in China. However, its impact on the biogeochemical conditions of sandstone uranium deposits remains poorly understood. In this study, we performed detailed physicochemical analyses of groundwater, as well as mineralogical, chemical and biogeochemical analyses of cores collected from the decommissioned AISL uranium mining area in the Yili Basin, northwestern China. After AISL uranium mining, U(IV) minerals and pyrite were oxidized and dissolved. The groundwater in the AISL-mined uranium deposit (AISLMUD) became highly acidic and oxidizing after mining ceased. Pyrite, which significantly influences the migration, transformation and immobilization of uranium, was minimal in all cores. Kaolinite, chlorite, quartz, K-feldspar, albite and muscovite exhibited poor adsorption capacity of U(VI) under strong acid conditions. The AISLMUD demonstrated limited capacity to retain U(VI) under highly acid conditions. Furthermore, high-throughput sequencing analysis revealed a substantial reduction in both microbial diversity and abundance in the cores. Microorganisms associated with U(VI) reduction were detected in the cores, with the exception of sulfate reduction bacteria. However, the results of microbial incubation showed that U(VI) reduction-related bacteria could not be in-situ activated by the addition of electron donors under the environmental stresses induced by prolonged acid in-situ leaching. These findings cast doubt on the feasibility of remediating uranium-contaminated groundwater with a pH below 3 through in-situ activation of U(VI) reduction-related bacteria. This research holds significant implications for evaluating the suitability of existing methods and developing new approaches for the restoration of AISLMUD.

Metagenomic survey reveals hydrocarbon biodegradation potential of Canadian high Arctic beaches

BackgroundDecreasing sea ice coverage across the Arctic Ocean due to climate change is expected to increase shipping activity through previously inaccessible shipping routes, including the Northwest Passage (NWP). Changing weather conditions typically encountered in the Arctic will still pose a risk for ships which could lead to an accident and the uncontrolled release of hydrocarbons onto NWP shorelines. We performed a metagenomic survey to characterize the microbial communities of various NWP shorelines and to determine whether there is a metabolic potential for hydrocarbon degradation in these microbiomes.ResultsWe observed taxonomic and functional gene evidence supporting the potential of NWP beach microbes to degrade various types of hydrocarbons. The metagenomic and metagenome-assembled genome (MAG) taxonomy showed that known hydrocarbon-degrading taxa are present in these beaches. Additionally, we detected the presence of biomarker genes of aerobic and anaerobic degradation pathways of alkane and aromatic hydrocarbons along with complete degradation pathways for aerobic alkane degradation. Alkane degradation genes were present in all samples and were also more abundant (33.8 ± 34.5 hits per million genes, HPM) than their aromatic hydrocarbon counterparts (11.7 ± 12.3 HPM). Due to the ubiquity of MAGs from the genus Rhodococcus (23.8% of the MAGs), we compared our MAGs with Rhodococcus genomes from NWP isolates obtained using hydrocarbons as the carbon source to corroborate our results and to develop a pangenome of Arctic Rhodococcus. Our analysis revealed that the biodegradation of alkanes is part of the core pangenome of this genus. We also detected nitrogen and sulfur pathways as additional energy sources and electron donors as well as carbon pathways providing alternative carbon sources. These pathways occur in the absence of hydrocarbons allowing microbes to survive in these nutrient-poor beaches.ConclusionsOur metagenomic analyses detected the genetic potential for hydrocarbon biodegradation in these NWP shoreline microbiomes. Alkane metabolism was the most prevalent type of hydrocarbon degradation observed in these tidal beach ecosystems. Our results indicate that bioremediation could be used as a cleanup strategy, but the addition of adequate amounts of N and P fertilizers, should be considered to help bacteria overcome the oligotrophic nature of NWP shorelines.

Ferrous-driven efficient removal of nitrate and various heavy metals by Zoogloea sp. ZP7 under oligotrophic conditions: Kinetics and heavy metal stress responses

Denitrification in surface water is affected by its oligotrophic characteristics and heavy metal pollution. In this study, a heterotrophic denitrifying bacterium Zoogloea resiniphila ZP7, which can utilize ferrous (Fe2+) as an additional electron donor under oligotrophic conditions, was isolated. Strain ZP7 achieved a 98.5 % nitrate (NO3−-N) removal efficiency (NRE) within 12 h using sodium acetate as carbon sources, with a Fe2+ concentration of 10.0 mg L−1, pH of 7.0, carbon to nitrogen ratio of 2.0, and NO3−-N concentration of 5.01 mg L−1, and no nitrite residue was observed. The denitrification performance and the removing ability of heavy metal by strain ZP7 were studied when exposed to copper (Cu2+), zinc (Zn2+), and cadmium (Cd2+) alone or in combination. When 1.0 mg L−1 Cu2+, Zn2+, and Cd2+ were added simultaneously, the strain ZP7 could achieve 91.3 % NRE in 14 h, and the removal efficiencies of Cu2+, Zn2+, and Cd2+ were 68.1, 89.2, and 83.5 %, respectively. The results of fluorescence region integration showed that the response of strain ZP7 to different heavy metal system stresses was different. Moreover, the changes in the content of extracellular polymeric substances (EPS) and humic acids indicated that they contributed to the enhancement of the resistance of strain ZP7 to heavy metals. Various characterization results showed that EPS and bio‑iron oxides were beneficial to remove heavy metals. In future studies, strain ZP7 can be considered to be immobilized in biomaterials to treat contaminated oligotrophic water bodies to explore its application prospects in actual contaminated water bodies.

Comparative analysis of sulfur-driven autotrophic denitrification for pilot-scale application: Pollutant removal performance and metagenomic function

Two parallel pilot-scale reactors were operated to investigate pollutant removal performance and metabolic pathways in elemental sulfur-driven autotrophic denitrification (SDAD) process under low temperature and after addition of external electron donors. The results showed that low temperature slightly inhibited SDAD (average total nitrogen removal of ∼4.7 mg L−1) while supplement of sodium thiosulfate (stage 2) and sodium acetate (stage 3) enhanced denitrification and secretion of extracellular polymeric substances (EPS), leading to the average removal rate of 0.75 and 1.01 kg N m−3 d−1, respectively with over twice higher total EPS. Correspondingly, nitrogen and sulfur related microbial metabolisms especially nitrite reductase and nitric oxide reductase encoding were promoted by genera including Thermomonas and Thiobacillus. The variations revealed that extra sodium acetate improved denitrification and enriched more SDAD-related microorganisms compared with sodium thiosulfate, which potentially catalyzed the refinement of practical strategies for optimizing denitrification in low carbon to nitrogen ratio wastewater treatment.

Activation of denitrification potential within anammox consortia to exceed nitrogen removal thresholds via Fe-Mn functionalized biochar: Enhanced interspecies electron transfer and metabolite cross-feeding

Anammox exerts a crucial role in nitrogenous effluents remediation; nevertheless, nitrogen removal efficiency thresholds triggered by additional nitrate production poses a critical obstacle for anammox performance. Herein, Fe-Mn functionalized biochar (FeMn#BC) was synthesized and incorporated into anammox consortia to elicit the exogenous organics-independent metabolic potential of denitrifying bacteria, which decreased nitrate accumulation and achieved up to 90.69 ± 0.48 % total nitrogen removal efficiency. Enzyme content analysis indicated that FeMn#BC bolstered biometabolic vigour and electron transport system activity by stimulating the synthesis of heme c and NADH. Microbial community succession results demonstrated that FeMn#BC facilitated the proliferation of anammox bacteria (AnAOB) and nitrate-reducing functional bacteria. Metagenomic analysis uncovered that FeMn#BC increased the relative abundance of genes related to energy metabolism, nutrient-substrate ingestion, cofactor synthesis, and iron-manganese transport pathways. Candidatus Brocadia sp013360945 interacted nitrogen metabolism substrates and electrons with dominated denitrifying species PR03 sp. (dependent on endogenous organics and FeMn#BC-released Fe(II)), while interacted cofactors with other mutualistic bacteria possessing nitrate-reducing potential (e.g. ATM1 sp003577165). FeMn#BC-released Mn(II) served as an additional electron donor, augmenting electron availability for non-dominant anammox species like Candidatus Jettenia caeni and Candidatus. Kuenenia stuttgartiensis. This research highlights the critical mechanism of Fe-Mn synergism for overcoming the bottleneck of anammox nitrogen removal efficiency, and demonstrates the substantial potential of Fe-Mn functionalized materials to revolutionize the existing wastewater treatment technologies towards a carbon-free approach.

Transparent insulating photochromic PU/PTA films for Wide-Spectrum modulated smart windows

Photochromic materials have recently attracted widespread attention in the research of smart windows because they can function without additional devices. In this study, an innovative photochromic thermal insulating hybrid film based on a polyurethane matrix was prepared via a one-pot method, incorporating phosphotungstic acid. The film demonstrates excellent transparency, good mechanical strength, and a significant modulation of the solar spectrum, transitioning from colorless to dark blue under sunlight without the need for additional electron donors. This photochromic behavior is reversible and reproducible without degradation, resulting in a transmittance variation of up to 71 % at 550 nm, which is the most sensitive to the human eye, and more than 50 % modulating performance in 1800 ∼ 2500 nm. In addition, thermal insulation experiments proved that the smart window device prepared by the film could achieve a reduction of 9.3 °C under daylight. Overall, the photochromic films prepared using the one-pot method show good performance involving mechanical strength, efficient photochromic response, broad spectral range modulation, high coloring efficiency and cycling stability, and have promising applications in energy-saving buildings, anti-counterfeiting and smart displays.

Anaerobic cyanides oxidation with bimetallic modulation of biological toxicity and activity for nitrite reduction

Cyanide is a typical toxic reducing agent prevailing in wastewater with a well-defined chemical mechanism, whereas its exploitation as an electron donor by microorganisms is currently understudied. Given that conventional denitrification requires additional electron donors, the cyanide and nitrogen can be eliminated simultaneously if the reducing HCN/CN− and its complexes are used as inorganic electron donors. Hence, this paper proposes anaerobic cyanides oxidation for nitrite reduction, whereby the biological toxicity and activity of cyanides are modulated by bimetallics. Performance tests illustrated that low toxicity equivalents of iron-copper composite cyanides provided higher denitrification loads with the release of cyanide ions and electrons from the complex structure by the bimetal. Both isotopic labeling and Density Functional Theory (DFT) demonstrated that CN−-N supplied electrons for nitrite reduction. The superposition of chemical processes reduces the biotoxicity and enhances the biological activity of cyanides in the CN−/Fe3+/Cu2+/NO2– coexistence system, including complex detoxification of CN− by Fe3+, CN− release by Cu2+ from [Fe(CN)6]3−, and NO release by nitrite substitution of −CN groups. Cyanide is the smallest structural unit of C/N-containing compounds and serves as a probe to extend the electron-donating principle of anaerobic cyanides oxidation to more electron-donor microbial utilization.

Nitrogen and sulfur cycling and their coupling mechanisms in eutrophic lake sediment microbiomes

Microorganisms play important roles in the biogeochemical cycles of lake sediment. However, the integrated metabolic mechanisms governing nitrogen (N) and sulfur (S) cycling in eutrophic lakes remain poorly understood. Here, metagenomic analysis of field and bioreactor enriched sediment samples from a typical eutrophic lake were applied to elucidate the metabolic coupling of N and S cycling. Our results showed significant diverse genes involved in the pathways of dissimilatory sulfur metabolism, denitrification and dissimilatory nitrate reduction to ammonium (DNRA). The N and S associated functional genes and microbial groups generally showed significant correlation with the concentrations of NH4+, NO2− and SO42, while with relatively low effects from other environmental factors. The gene-based co-occurrence network indicated clear cooperative interactions between N and S cycling in the sediment. Additionally, our analysis identified key metabolic processes, including the coupled dissimilatory sulfur oxidation (DSO) and DNRA as well as the association of thiosulfate oxidation complex (SOX systems) with denitrification pathway. However, the enriched N removal microorganisms in the bioreactor ecosystem demonstrated an additional electron donor, incorporating both the SOX systems and DSO processes. Metagenome-assembled genomes-based ecological model indicated that carbohydrate metabolism is the key linking factor for the coupling of N and S cycling. Our findings uncover the coupling mechanisms of microbial N and S metabolism, involving both inorganic and organic respiration pathways in lake sediment. This study will enhance our understanding of coupled biogeochemical cycles in lake ecosystems.

Biostimulation with oxygen and electron donors supports micropollutant biodegradation in an experimentally simulated nitrate-reducing aquifer

The availability of suitable electron donors and acceptors limits micropollutant natural attenuation in oligotrophic groundwater. This study investigated how electron donors with different biodegradability (humics, dextran, acetate, and ammonium), and different oxygen concentrations affect the biodegradation of 15 micropollutants (initial concentration of each micropollutant = 50 μg/L) in simulated nitrate reducing aquifers. Tests mimicking nitrate reducing field conditions showed no micropollutant biodegradation, even with electron donor amendment. However, 2,4-dichlorophenoxyacetic acid and mecoprop were biodegraded under (micro)aerobic conditions with and without electron donor addition. The highest 2,4-dichlorophenoxyacetic acid and mecoprop biodegradation rates and removal efficiencies were obtained under fully aerobic conditions with amendment of an easily biodegradable electron donor. Under microaerobic conditions, however, amendment with easily biodegradable dissolved organic carbon (DOC) inhibited micropollutant biodegradation due to competition between micropollutants and DOC for the limited oxygen available. Microbial community composition was dictated by electron acceptor availability and electron donor amendment, not by micropollutant biodegradation. Low microbial community richness and diversity led to the absence of biodegradation of the other 13 micropollutants (such as bentazon, chloridazon, and carbamazepine). Finally, adaptation and potential growth of biofilms interactively determined the location of the micropollutant removal zone relative to the point of amendment. This study provides new insight on how to stimulate in situ micropollutant biodegradation to remediate oligotrophic groundwaters as well as possible limitations of this process.

Response mechanism of microalgae-based constructed wetland to day-night alternations

As an effective optimization strategy for constructed wetlands, microalgae-constructed wetland coupling systems (MCCSs) carry the potential risk of eutrophication during operation. Within MCCSs, the key microorganisms that trigger eutrophication are highly responsive to day-night alternations. Hence, optimizing the day-to-night ratio may be an effective measure to reduce the eutrophication risk from MCCSs. To verify this hypothesis, we analyzed the response characteristics of MCCSs under different day-night cycles (light/dark: 9 h/15 h, 12 h/12 h, 15 h/9h, and light 24 h). The results indicate that in response to day-night changes, the microbial communities in the MCCSs established stable microbial networks through competitive interactions. Furthermore, energy metabolism played a crucial role in the influence of the day-night alternations on the microbial functional metabolism in the MCCSs. Moderate extension of light time can provide additional electron donors for microbial denitrification, thereby maintaining the stability of denitrification while increasing the dissolved oxygen concentration. The results of the structural equation model also confirmed that the microbial communities contributed more to the environmental function of the MCCSs through energy metabolism under light conditions. This study provides valuable insights into the prevention of eutrophication and maintenance of stable operation in MCCSs.

Enhanced nitrate removal efficiency and microbial response of immobilized mixed aerobic denitrifying bacteria through biochar coupled with inorganic electron donors in oligotrophic water

The nitrogen removal characteristics and microbial response of biochar-immobilized mixed aerobic denitrifying bacteria (BIADB) were investigated at 25 °C and 10 °C. BIADB removed 53.51 ± 1.72 % (25 °C) and 39.90 ± 4.28 % (10 °C) nitrate in synthetic oligotrophic water. Even with practical oligotrophic water, BIADB still effectively removed 47.66–53.21 % (25 °C), and 39.26–45.63 % (10 °C) nitrate. The addition of inorganic electron donors increased nitrate removal by approximately 20 % for synthetic and practical water. Bacterial and functional communities exhibited significant temperature and stage differences (P < 0.05), with temperature and total dissolved nitrogen being the main environmental factors. The dominant genera and keystone taxa exhibited significant differences at the two temperatures. Structural equation model analysis showed that dissolved organic matter had the highest direct and indirect effects on diversity and function, respectively. This study provides an innovative pathway for utilizing biochar and inorganic electron donors for nitrate removal from oligotrophic waters.

Acetic acid stress response of the acidophilic sulfate reducer Acididesulfobacillus acetoxydans.

Acid mine drainage (AMD) waters are a severe environmental threat, due to their high metal content and low pH (pH <3). Current technologies treating AMD utilize neutrophilic sulfate-reducing microorganisms (SRMs), but acidophilic SRM could offer advantages. As AMDs are low in organics these processes require electron donor addition, which is often incompletely oxidized into organic acids (e.g., acetic acid). At low pH, acetic acid is undissociated and toxic to microorganisms. We investigated the stress response of the acetotrophic Acididesulfobacillus acetoxydans to acetic acid. A. acetoxydans was cultivated in bioreactors at pH 5.0 (optimum). For stress experiments, triplicate reactors were spiked until 7.5 mM of acetic acid and compared with (non-spiked) triplicate reactors for physiological, transcriptomic, and membrane lipid changes. After acetic acid spiking, the optical density initially dropped, followed by an adaptation phase during which growth resumed at a lower growth rate. Transcriptome analysis revealed a downregulation of genes involved in glutamate and aspartate synthesis following spiking. Membrane lipid analysis revealed a decrease in iso and anteiso fatty acid relative abundance; and an increase of acetyl-CoA as a fatty acid precursor. These adaptations allow A. acetoxydans to detoxify acetic acid, creating milder conditions for other microorganisms in AMD environments.

Metabolite cross-feeding enables concomitant catabolism of chlorinated methanes and chlorinated ethenes in synthetic microbial assemblies.

Isolate studies have been a cornerstone for unraveling metabolic pathways and phenotypical (functional) features. Biogeochemical processes in natural and engineered ecosystems are generally performed by more than a single microbe and often rely on mutualistic interactions. We demonstrate the rational bottom-up design of synthetic, interdependent co-cultures to achieve concomitant utilization of chlorinated methanes as electron donors and organohalogens as electron acceptors. Specialized anaerobes conserve energy from the catabolic conversion of chloromethane or dichloromethane to formate, H2, and acetate, compounds that the organohalide-respiring bacterium Dehalogenimonas etheniformans strain GP requires to utilize cis-1,2-dichloroethenene and vinyl chloride as electron acceptors. Organism-specific qPCR enumeration matched the growth of individual dechlorinators to the respective functional (i.e. dechlorination) traits. The metabolite cross-feeding in the synthetic (co-)cultures enables concomitant utilization of chlorinated methanes (i.e. chloromethane and dichloromethane) and chlorinated ethenes (i.e. cis-1,2-dichloroethenene and vinyl chloride) without the addition of an external electron donor (i.e. formate and H2). The findings illustrate that naturally occurring chlorinated C1 compounds can sustain anaerobic food webs, an observation with implications for the development of interdependent, mutualistic communities, the sustenance of microbial life in oligotrophic and energy-deprived environments, and the fate of chloromethane/dichloromethane and chlorinated electron acceptors (e.g. chlorinated ethenes) in pristine environments and commingled contaminant plumes.

A Multifunctional Dehalobacter? Tandem Chloroform and Dichloromethane Degradation in a Mixed Microbial Culture.

Chloroform (CF) and dichloromethane (DCM) contaminate groundwater sites around the world but can be cleaned up through bioremediation. Although several strains of Dehalobacter restrictus can reduce CF to DCM and multiple Peptococcaceae can ferment DCM, these processes cannot typically happen simultaneously due to CF sensitivity in the known DCM-degraders or electron donor competition. Here, we present a mixed microbial culture that can simultaneously metabolize CF and DCM and create an additional enrichment culture fed only DCM. Through genus-specific quantitative polymerase chain reaction, we find that Dehalobacter grows while either CF alone or DCM alone is converted, indicating its involvement in both metabolic steps. Additionally, the culture was maintained for over 1400 days without the addition of an exogenous electron donor, and through electron balance calculations, we show that DCM metabolism would produce sufficient reducing equivalents (likely hydrogen) for CF respiration. Together, these results suggest intraspecies electron transfer could occur to continually reduce CF in the culture. Minimizing the addition of electron donor reduces the cost of bioremediation, and "self-feeding" could prolong bioremediation activity long after donor addition ends. Overall, understanding this mechanism informs strategies for culture maintenance and scale-up and benefits contaminated sites where the culture is employed for remediation worldwide.

Pore-Confined π-Chromophoric Tetracene as a Visible Light Harvester toward MOF-Based Photocatalytic CO2 Reduction in Water.

Integrating photoactive π-chromophoric guest molecules inside the MOF nanopore can result in the emergence of light-responsive features, which in turn can be utilized for developing photoactive materials with inherent properties of MOF. Herein, we report the confining of π-chromophoric tetracene (TET) molecules inside the nanospace of postmodified Zr-MOF-808 (Zr-MOF) with MBA molecules (MBA = 2-(5'-methyl-[2,2'-bipyridine]-5-yl)acetic acid) for effectively utilizing its light-harvesting properties toward photocatalytic CO2 reduction. The confinement of the TET molecules as a photosensitizer and the covalent grafting of a catalytically active [Re(MBA)(CO)3Cl] complex, postsynthetically, result in a single integrated catalytic system named Zr-MBA-TET-Re-MOF. Photoreduction of CO2 over Zr-MBA-TET-Re-MOF showed the evolution of 805 μmol g-1 CO with 99.9% selectivity after 10 h of continuous visible light irradiation in water without any additional sacrificial electron donor and having the apparent quantum efficiency of 1.3%. In addition, the catalyst demonstrated an appreciable activity even under direct sunlight irradiation in aqueous medium with a maximum production of 362.7 μmol g-1 CO, thereby mimicking artificial photosynthesis. Moreover, electron transfer from TET to the catalytic center was supported by the formation of photoinduced TET radical cation, as inferred from in situ UV-vis spectra, electron paramagnetic resonance (EPR) analysis, and transient absorption (TA) studies. Additionally, the in situ diffuse reflectance infrared Fourier transform (DRIFT) measurements support that the photoreduction of CO2 to CO proceeds via *COOH intermediate formation. The close proximity of the light-harvesting molecule and catalytic center facilitated facile electron transfer from the photosensitizer to the catalyst during the CO2 reduction.

Long-term effects of thiosulfate on the competition between sulfur-mediated bacteria and glycogen accumulating organisms in sulfate-rich carbon-deficient wastewater

Sewage nutrient (e.g., nitrogen and phosphorus) biological removal performance is often limited by the deficient carbon source and undesirable glycogen accumulating organisms (GAOs), even in sulfate-containing wastewater. Thiosulfate (S2O32−) as a bioavailable, environmentally-benign, metastable and cost-effective agent has been regarded as electron carriers that induces high sulfur-mediated bacterial activity for nutrient removal from wastewater. In this study, the long-term effects of thiosulfate on the competition between sulfur-mediated bacteria (SMB, including sulfur-reducing bacteria (SRB) and sulfur-oxidizing bacteria (SOB)) and GAOs were explored to further close the gap of our knowledge on the control of GAOs under carbon deficient wastewater. Three reactors were continuously operated for over 100 days and were fed with 200 mg acetate-COD/L and 20 (R1), 50 (R2) and 80 (R3) mg S/L thiosulfate respectively. The results revealed that adding thiosulfate at the beginning of the anoxic phase promoted sulfur metabolism and increased the proliferation of SRB (mainly Desulfobacter) and SOB (mainly Chromatiaceae). Correspondingly, the relative abundance of GAOs (mainly Candidatus_Competibacter) decreased. After the carbon source was reduced, the abundance of GAOs increased and the competitive activity of SRB was weakened, resulting in the reduced sulfate reduction, which could be attributed to the fact that GAOs had a higher carbon source competitiveness than SRB under low carbon source conditions. While SOB maintained a high abundance due to the addition of thiosulfate as an additional electron donor, which enhanced the denitrification efficiency. Additionally, the dominant SOB shifted from Thiobacillus to Chromatiaceae during the long-term operation, indicating that Chromatiaceae had a higher competitive advantage for reduced sulfur (e.g., S2O32−, Polysulfide (Poly-S)) and nitrate compared to Thiobacillus. Furthermore, microbial functional genes revealed that S metabolism was enhanced during long-term operation. The potential mechanism and optimization strategy regarding the competition between sulfur-mediated bacteria and GAOs were revealed.