Nitrogenases are the only known enzymes that reduce molecular nitrogen (N2) to ammonia. Recent findings have demonstrated that nitrogenases also reduce the greenhouse gas carbon dioxide (CO2), suggesting CO2 to be a competitor of N2. However, the impact of omnipresent CO2 on N2 fixation has not been investigated to date. Here, we study the competing reduction of CO2 and N2 by the two nitrogenases of Rhodobacter capsulatus, the molybdenum and the iron nitrogenase. The iron nitrogenase is almost threefold more efficient in CO2 reduction and profoundly less selective for N2 than the molybdenum isoform under mixtures of N2 and CO2. Correspondingly, the growth rate of diazotrophically grown R. capsulatus strains relying on the iron nitrogenase notably decreased after adding CO2. The in vivo CO2 activity of the iron nitrogenase facilitates the light-driven extracellular accumulation of formate and methane, one-carbon substrates for other microbes, and feedstock chemicals for a circular economy.

The thermodynamic effect of octyl-β-D-glucopyranoside (OGP) on the formation of methane-1,3-dimethylcyclohexane (DMCH) hydrate was studied in this work. The thermodynamic equilibrium hydrate formation pressures between 275.15 K and 283.15 K were measured by the isothermal pressure search method. Different OGP aqueous solutions (0, 0.1, and 1 wt%) were used in this work. The experimental results show that OGP had no obvious thermodynamic inhibition on methane-DMCH hydrate formation when its concentration was low (0.1 wt%), whereas it had an inhibition on methane-DMCH hydrate formation when its concentration was high (1 wt%). The phase equilibrium hydrate formation pressure of the methane-DMCH-OGP system is about 0.1 MPa higher than that of the methane-DMCH system. The dissociation enthalpies of methane hydrate in different solutions remained uniform, which indicates that OGP was not involved in methane-DMCH hydrate formation. This phenomenon is explained from the perspective of the molecular structure of OGP. As a renewable and biological nonionic surfactant, the concentration of OGP in the liquid phase is low, so OGP can be added to the methane-DMCH system without significant thermodynamic inhibition.

High-level erythromycin (ERY) fermentation wastewater will pose serious threats to lake environments. Anaerobic digestion (AD) has advantages in treating high-level antibiotic wastewater. However, the fate of antibiotic resistance genes (ARGs) and microbial communities in AD after stepwise exposure to high-level ERY remains unclear. In this study, an AD reactor was first exposed to 0, 5, 10, 50, 100 and 200 mg/L ERY and then re-exposed to 0, 50, 200 and 500 mg/L ERY to investigate the effect of ERY on AD. The results show that AD could adapt to the presence of high-level ERY (500 mg/L) and could maintain efficient CH4 production after domestication with low-level ERY (50 mg/L). The AD process could achieve higher removal of ERY (>94%), regardless of the initial ERY concentration. ErmB and mefA, conferring resistance through target alteration and efflux pumps, respectively, were dominant in the AD process. The first exposure to ERY stimulated an increase in the total ARG abundance, while the AD process seemed to discourage ARG maintenance following re-exposure to ERY. ERY inhibited the process of acetoclastic methanogenesis, but strengthened the process of hydrogenotrophic methanogenesis. This work provides useful information for treating high-level ERY fermentation wastewater by the AD process.

Fast, accurate, real-time measurement of gas concentration is an important task for preventing coal mining disasters. In order to develop an accurate monitoring method for methane gas concentration at different locations in a mine environment, a non-source optical fiber sensor for multi-point methane detection has been developed in this paper. A 16-channel fiber splitter and a multi-channel time-sharing acquisition module are employed within the sensor, enabling simultaneous detection of methane gas at 16 points by a host. Furthermore, the methane sensors are connected to the monitoring host via an all-optical method, achieving non-source and long-range detection of methane. To assess the performance of the methane gas sensor, experiments were conducted to evaluate its detection range, response time, and stability. The results indicated that the average detection error was approximately 1.84% across the full range, and the response time did not exceed 10 s. The minimum detection limit of the sensor, as determined by the 1σ criteria, was obtained as 58.42 ppm. Additionally, the concentrations of methane gas measured at varying distances (1 km, 2 km, 5 km) were found to be essentially consistent over an extended period. These results suggest that the development of this non-source optical fiber sensor holds significant potential for providing a method for mine environment, multi-point online methane gas detection.

This study evaluates the effects of substituting cassava pulp with broken rice and cassava chips in the total mixed ration silage diets of beef cattle on feed composition, ensiling quality, digestibility, and energy utilization. Fifteen Holstein Thai native crossbred (89% Bos taurus × 11% Bos indicus) steers in the fattening phase, with an average age of 2.5 ± 0.1 years and an initial body weight of 603.7 ± 14.3 kg, were used in the energy balance trial. Using a randomized complete block design with five replications, the steers received one of three treatments. The three dietary treatments included substituting cassava pulp with cassava chips and broken rice on a dry matter basis with ratios of 50:0:0, 30:20:0, or 10:20:20. The results show that broken rice is a superior nutrient source and provides greater energy balance (p < 0.01). Despite the cost implications, substituting cassava pulp and chips positively impacts the ensilage pH and reduces the acetic acid concentration (p < 0.01). There was an increase in the lactic acid bacteria count (p < 0.05) and a reduction in the rumen ammonia, propionate, and butyrate concentrations (p < 0.05) without adverse effects (p > 0.05) on digestibility, blood metabolites, or enteric methane emissions. These findings suggest that broken rice is a promising alternative grain-rich ruminant feed. Future research should explore on-farm long-term feeding and economic evaluations to provide a more comprehensive understanding of the practical implications.

Reducing methane (CH4) emissions is increasingly recognized as an urgent greenhouse gas mitigation priority for avoiding ecosystem \'tipping points\' that will accelerate global warming. Agricultural systems, namely ruminant livestock and rice cultivation are dominant sources of CH4 emissions. Efforts to reduce methane from rice typically focus on water management strategies that implicitly assume that irrigated rice systems are consistently flooded and that farmers exert a high level of control over the field water balance. In India most rice is cultivated during the monsoon season and hydrologic variability is common, particularly in the Eastern Gangetic Plains (EGP) where high but variable rainfall, shallow groundwater, and subtle differences in topography interact to create complex mosaics of field water conditions. Here, we characterize the hydrologic variability of monsoon season rice fields (n = 207) in the Indian EGP (\'Eastern India\') across two contrasting climate years (2021, 2022) and use the Denitrification Decomposition (DNDC) model to estimate GHG emissions for the observed hydrologic conditions. Five distinct clusters of field hydrology patterns were evident in each year, but cluster characteristics were not stable across years. In 2021, average GHG emissions (8.14 mt CO2-eq ha-1) were twice as high as in 2022 (3.81 mt CO2-eq ha-1). Importantly, intra-annual variability between fields was also high, underlining the need to characterize representative emission distributions across the landscape and across seasons to appropriately target GHG mitigation strategies and generate accurate baseline values. Simulation results were also analyzed to identify main drivers of emissions, with readily identified factors such as flooding period and hydrologic interactions with crop residues and nitrogen management practices emerging as important. These insights provide a foundation for understanding landscape variability in GHG emissions from rice in Eastern India and suggest priorities for mitigation that honor the hydrologic complexity of the region.

Anaerobic digestion (AD) technology can potentially address the gap between energy demand and supply playing a crucial role in the production of sustainable energy from utilization of biogenic waste materials as feedstock. The biogas production from anaerobic digestion is primarily influenced by the chemical compositions and biodegradability of the feedstock. Organosolv-steam explosion offers a constructive approach as a promising pretreatment method for the fractionation of lignocellulosic biomasses delivering high cellulose content.This study showed how synergetic co-digestion serves to overcome the challenges of mono-digestion\'s low efficiency. Particularly, the study evaluated the digestibility of organosolv-steam pretreated wheat straw (WSOSOL) in mono as well as co-digesting substrate with cheese whey (CW) and brewery spent grains (BSG). The highest methane yield was attained with co-digestion of WSOSOL + CW (338 mL/gVS) representing an enhanced biogas output of 1-1.15 times greater than its mono digestion. An ammonium production was favored under co-digestion strategy accounting for 921 mg/L from WSOSOL + BSG. Metagenomic study was conducted to determine the predominant bacteria and archaea, as well as its variations in their populations and their functional contributions during the AD process. The Firmicutes have been identified as playing a significant role in the hydrolysis process and the initial stages of AD. An enrichment of the most prevalent archaea genera enriched were Methanobacterium, Methanothrix, and Methanosarsina. Reactors digesting simpler substrate CW followed the acetoclastic, while digesting more complex substrates like BSG and WSOSOL followed the hydrogenotrophic pathway for biomethane production. To regulate the process for an enhanced AD process to maximize CH4, a comprehensive understanding of microbial communities is beneficial.

High ammonia concentrations in anaerobic degradation systems cause volatile fatty acid accumulation and reduced methane yield, which often derive from restricted activity of syntrophic acid-oxidising bacteria and hydrogenotrophic methanogens. Inclusion of additives that facilitate the electron transfer or increase cell proximity of syntrophic species by flocculation can be a suitable strategy to counteract these problems, but its actual impact on syntrophic interactions has yet to be determined. In this study, microbial cultivation and molecular and microscopic analysis were performed to evaluate the impact of conductive (graphene, iron oxide) and non-conductive (zeolite) additives on the degradation rate of acetate and propionate to methane by highly enriched ammonia-tolerant syntrophic cultures derived from a biogas process. All additives had a low impact on the lag phase but resulted in a higher rate of acetate (except graphene) and propionate degradation. The syntrophic bacteria \'Candidatus Syntrophopropionicum ammoniitolerans\', Syntrophaceticus schinkii and a novel hydrogenotrophic methanogen were found in higher relative abundance and higher gene copy numbers in flocculating communities than in planktonic communities in the cultures, indicating benefits to syntrophs of living in close proximity to their cooperating partner. Microscopy and element analysis showed precipitation of phosphates and biofilm formation in all batches except on the graphene batches, possibly enhancing the rate of acetate and propionate degradation. Overall, the concordance of responses observed in both acetate- and propionate-fed cultures highlight the suitability of the addition of iron oxide or zeolites to enhance acid conversion to methane in high-ammonia biogas processes. KEY POINTS: • All additives promoted acetate (except graphene) and propionate degradation. • A preference for floc formation by ammonia-tolerant syntrophs was revealed. • Microbes colonised the surfaces of iron oxide and zeolite, but not graphene.

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BACKGROUND: Xanthenes and multi-aryl carbon core containing compounds represent different types of complex and condensed architectures that have impressive wide range of pharmacological, industrial and synthetic applications. Moreover, indoles as building blocks were only found in naturally occurring metabolites with di-aryl carbon cores and in chemically synthesized tri-aryl carbon core containing compounds. Up to date, rare xanthenes with indole bearing multicaryl carbon core have been reported in natural or synthetic products. The underlying mechanism of fluorescein-like arthrocolins with tetra-arylmethyl core were synthesized in an engineered Escherichia coli fed with toluquinol remained unclear.
RESULTS: In this study, the Keio collection of single gene knockout strains of 3901 mutants of E. coli BW25113, together with 14 distinct E. coli strains, was applied to explore the origins of endogenous building blocks and the biogenesis for arthrocolin assemblage. Deficiency in bacterial respiratory and aromatic compound degradation genes ubiX, cydB, sucA and ssuE inhibited the mutant growth fed with toluquinol. Metabolomics of the cultures of 3897 mutants revealed that only disruption of tnaA involving in transforming tryptophan to indole, resulted in absence of arthrocolins. Further media optimization, thermal cell killing and cell free analysis indicated that a non-enzyme reaction was involved in the arthrocolin biosynthesis in E. coli. Evaluation of redox potentials and free radicals suggested that an oxygen-mediated free radical reaction was responsible for arthrocolins formation in E. coli. Regulation of oxygen combined with distinct phenol derivatives as inducer, 31 arylmethyl core containing metabolites including 13 new and 8 biological active, were isolated and characterized. Among them, novel arthrocolins with p-hydroxylbenzene ring from tyrosine were achieved through large scale of aerobic fermentation and elucidated x-ray diffraction analysis. Moreover, most of the known compounds in this study were for the first time synthesized in a microbe instead of chemical synthesis. Through feeding the rat with toluquinol after colonizing the intestines of rat with E. coli, arthrocolins also appeared in the rat blood.
CONCLUSIONS: Our findings provide a mechanistic insight into in vivo synthesis of complex and condensed arthrocolins induced by simple phenols and exploits a quinol based method to generate endogenous aromatic building blocks, as well as a methylidene unit, for the bacteria-facilitated synthesis of multiarylmethanes.