Oxidation Of Hydroxylamine Research Articles

Ultrasound-assisted bisphenol AF degradation using in situ generated hydrogen peroxide

Bisphenol AF (BPAF) is degraded through the ultrasound-assisted in situ generation and activation of hydrogen peroxide (H2O2) by the copper(II) catalysed oxidation of hydroxylamine (NH2OH) with dioxygen (O2). Compared to added H2O2, in situ generated H2O2 significantly improves the degradation of BPAF from 46.7% to 94.8% in ∼15 min. The reaction follows a pseudo-first-order kinetic model. This study examines the influence of solution pH, anions, humic acid, and different concentrations of the reactants on BPAF degradation. Mass spectrometry was used to identify the BPAF degradation products, and a degradation pathway is proposed. This work advances the understanding of in situ hydrogen peroxide generation and activation in advanced oxidation (Fenton-like) processes (AOPs).

Co-composting of tail vegetable with flue-cured tobacco leaves: analysis of nitrogen transformation and estimation as a seed germination agent for halophyte.

Resource utilization of tail vegetables has raised increasing concerns in the modern agriculture. However, the effect and related mechanisms of flue-cured tobacco leaves on the product quality, phytotoxicity and bacterially-mediated nitrogen (N) transformation process of tail vegetable composting were poorly understood. Amendments of high-dosed (5% and 10% w/w) tobacco leaves into the compost accelerated the heating process, prolonged the time of thermophilic stage, increased the peak temperature, thereby improving maturity and shortening composting duration. The tobacco leaf amendments at the 10% (w/w) increased the N conservation (TN and NH4-N content) of compost, due to the supply of N-containing nutrient and promotion of organic matter degradation by tobacco leaves. Besides, tobacco leaf amendments promoted the seed germination and root development of wild soybean, exhibiting the feasibility of composting product for promoting the growth of salt-tolerant plants, but no dose-dependent effect was found for tobacco leaf amendments. Addition of high dosed (5% and 10% w/w) tobacco leaves shifted the bacterial community towards lignocellulosic and N-fixing bacteria, contributing to increasing the compost maturity and N retention. PICRUSt 2 functional prediction revealed that N-related bacterial metabolism (i.e., hydroxylamine oxidation and denitrifying process) was enhanced in the tobacco leaf treatments, which contributed to N retention and elevated nutrient quality of composting. To the best knowledge, this was the first study to explore the effect of tobacco waste additives on the nutrient transformation and halophyte growth promotion of organic waste composting. These findings will deepen the understanding of microbially-mediated N transformation and composting processes involving flue-cured tobacco leaves.

Nitrogen input modulates the effects of coastal acidification on nitrification and associated N2O emission

Acidification of coastal waters, synergistically driven by increasing atmospheric carbon dioxide (CO2) and intensive land-derived nutrient inputs, exerts significant stresses on the biogeochemical cycles of coastal ecosystem. However, the combined effects of anthropogenic nitrogen (N) inputs and aquatic acidification on nitrification, a critical process of N cycling, remains unclear in estuarine and coastal ecosystems. Here, we showed that increased loading of ammonium (NH4+) in estuarine and coastal waters alleviated the inhibitory effect of acidification on nitrification rates but intensified the production of the potent greenhouse gas nitrous oxide (N2O), thus accelerating global climate change. Metatranscriptomes and natural N2O isotopic signatures further suggested that the enhanced emission of N2O may mainly source from hydroxylamine (NH2OH) oxidation rather than from nitrite (NO2−) reduction pathway of nitrifying microbes. This study elucidates how anthropogenic N inputs regulate the effects of coastal acidification on nitrification and associated N2O emissions, thereby enhancing our ability to predict the feedbacks of estuarine and coastal ecosystems to climate change and human perturbations.

Integrative leaf anatomy structure, physiology, and metabolome analyses revealed the response to drought stress in sainfoin at the seedling stage.

Sainfoin (Onobrychis viciaefolia) is a vital legume forage, and drought is the primary element impeding sainfoin growth. The anatomical structure, physiological indexes, and metabolites of the leaves of sainfoin seedlings with a drought-resistant line of P1 (DRL) and a drought-sensitive material of 2049 (DSM) were analyzed under drought (-1.0 MPa) with polyethylene glycol-6000 (PEG-6000). The leaf anatomy was studied by the paraffin section method. The related physiological indexes were measured by the hydroxylamine oxidation method, titanium sulfate colorimetric method, thiobarbituric acid method, acidic ninhydrin colorimetric method, and Coomassie brilliant blue method. The metabolomics analysis was composed of liquid chromatography tandem high-resolution mass spectrometry (LC-MS/MS). The results revealed that the thickness of the epidermis, palisade tissue, and sponge tissue of DRL were significantly greater than those of DSM. The leaves of DRL exhibited lower levels of superoxide anion (O2 •-) production rate, hydrogen peroxide (H2O2) content, and malondialdehyde (MDA) content compared with DSM, while proline (Pro) content and soluble protein (SP) content were significantly higher than those of DSM. A total of 391 differential metabolites were identified in two samples. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment showed that the primary differential metabolites were concentrated into the tyrosine metabolism; isoquinoline alkaloid biosynthesis; ubiquinone and other terpenoid quinone biosynthesis; neomycin, kanamycin, and gentamicin biosynthesis; and anthocyanin biosynthesis metabolic pathways. Compared with DSM, DRL had more complete anatomical structure, lower active oxygen content, and higher antioxidant level. The results improved our insights into the drought-resistant mechanisms in sainfoin.

Micro-oxygen supply combined with applied voltage enhance ammonium oxidation and denitrification in a single-chamber microbial electrolysis cell: Performance and mechanism

Anodic ammonium oxidation catalyzed by microorganisms holds great potential as an effective anaerobic approach for ammonium and total nitrogen removal. However, the application of this approach is constrained by a low rate of ammonium oxidation, limiting the efficiency of total nitrogen removal. To overcome this limitation, a small amount of oxygen was introduced into a single-chamber microbial electrolysis cell to initiate the production of hydroxylamine through ammonium oxidation. Enhanced efficiency of overall nitrogen removal was obtained by the combined regulation of micro-oxygen and microbial electrode catalysis for ammonium oxidation and denitrification. According to the ammonium oxidation amount, the total oxidized nitrogen removal efficiency reached 67.87 ± 0.25 % and total nitrogen removal amount was 202.9 ± 0.5 mg/L, showing a high performance for treating high ammonium-contained wastewater. The oxidation peaks of hydroxylamine were determined at 0.43 V (vs. Ag/AgCl) for revealing the possible process of anodic hydroxylamine oxidation coupling to cathodic denitrification. In addition, microbial community analysis revealed that increasing the applied voltage regulated the abundance of nitrifying and denitrifying functional microbial communities both in the solution and on electrodes, and the key functional genes including hydroxylamine oxidation (hao), nitrate reduction (narG, narH, narI, napA, napB), nitrite reduce (nirK, nirS), and nitric oxide reduce (norB, norC), increased significantly at the applied voltage of 0.9 V, which accompanied by an obvious enhancement of enzyme activity compared to anaerobic and open circuit voltage. This work revealed the enhanced performance and mechanism on synergistic regulation by micro-oxygen and applied voltage for ammonium oxidation and denitrification, and provided new insights and feasible enhancing measures for treating high ammonium-nitrogen wastewater with low C/N ratio.

Polyvinyl Chloride Microplastics Facilitate Nitrous Oxide Production in Partial Nitritation Systems.

Partial nitritation (PN) is an important partner with anammox in the sidestream line treating high-strength wastewater and primarily contributes to nitrous oxide (N2O) emissions in such a hybrid system, which also suffers from ubiquitous microplastics because of the growing usage and disposal levels of plastics. In this study, the influences of polyvinyl chloride microplastics (PVC-MPs) on N2O-contributing pathways were experimentally revealed to fill the knowledge gap on N2O emission from the PN system under microplastics stress. The long-term results showed that the overall PN performance was hardly affected by the low-dose PVC-MPs (0.5 mg/L) while obviously deteriorated by the high dose (5 mg/L). According to the batch tests, PVC-MPs reduced biomass-specific ammonia oxidation rates (AORs) by 5.78-21.94% and stimulated aerobic N2O production by 9.22-88.36%. Further, upon increasing dissolved oxygen concentrations from 0.3 to 0.9 mg O2/L, the degree of AOR inhibition increased but that of N2O stimulation was lightened. Site preference analysis in combination with metabolic inhibitors demonstrated that the contributions of hydroxylamine oxidation and heterotrophic denitrification to N2O production at 0.3 mg O2/L were enhanced by 18.84 and 10.34%, respectively, accompanied by a corresponding decreased contribution of nitrifier denitrification. Finally, the underlying mechanisms proposed for negative influences of PVC-MPs were bisphenol A leaching and reactive oxygen species production, which led to more cell death, altered sludge properties, and reshaped microbial communities, further resulting in enhanced N2O emission. Overall, this work implied that the ubiquitous microplastics are a hidden danger that cannot be ignored in the PN system.

Genome-resolved metagenomics provides insights into the microbial-mediated sulfur and nitrogen cycling in temperate seagrass meadows

The presence of seagrasses facilitates numerous microbial-mediated biogeochemical cycles, with sulfur- and nitrogen-cycling microorganisms playing crucial roles as regulators. Despite efforts to comprehend the diversity of microbes in seagrass ecosystems, the metabolic functions of these benthic microorganisms in seagrass sediments remain largely unknown. Using metagenomics, we provide insights into the sulfur- and nitrogen-cycling pathways and key metabolic capacities of microorganisms in both Z. japonica-colonized and unvegetated sediments over a seasonal period. Taxonomic analysis of N and S cycling genes revealed that δ- and γ- proteobacteria dominated the benthic sulfate-reducing bacteria, while α- and γ-proteobacteria played a significant role in the sulfur-oxidation processes. The proteobacterial lineages were also major contributors to the benthic nitrogen cycling. However, at a finer taxonomic resolution, microbial participants in different processes were observed to be highly diverse and mainly driven by environmental factors such as temperature and salinity. The gene pools of sulfur and nitrogen cycles in the seagrass sediments were dominated by genes involved in sulfide oxidation (fccA) and hydroxylamine oxidation (hao), respectively. Seagrass colonization elevated the relative abundance of genes responsible for sulfite production (phsC), hydroxylamine oxidation (hao), and nitrogen fixation (nifK), but suppressed sulfur oxidation (soxXYZ) and denitrification (nosZ and nirS). The prevalence of proteobacterial lineages functioned with versatile capabilities in both sulfur and nitrogen cycles in seagrass ecosystems, highlighting tight couplings between these processes, which was further supported by the recovery of 83 metagenome-assembled genomes (MAGs). These findings broaden our understanding of the biogeochemical processes that are mediated by microorganisms in seagrass ecosystems.

Allosteric switch for electrochemical aptasensor toward heavy metals pollution of Lentinus edodes sensitized with porphyrinic metal-organic frameworks

BackgroundLentinan medicament from Lentinus edodes has been considered as natural medicinal products with minimal side effects for cancer therapy, but Lentinus edodes are easily polluted by nonbiodegradable heavy metals, especially silver ion (Ag+). Therefore, it is highly desirable to monitor Ag + pollution in Lentinus edodes considering their adverse impact on lentinan medicament. Electrochemical sensor isn't affected from the interference of matrix turbidity and color, and offers a powerful means for determination of variant analytes. As for electrochemical sensing toward Ag+, there is a great need to design efficient signal probes for specific recognition and signal generation. ResultsWe present an appropriate electrochemical aptasensor for Ag + assay based on biomimetic catalysis of porphyrin-encapsulated MOF (PorMOF) and allosteric switch of C-rich DNA. Thanks to the excellent biocompatibility, PorMOFs as nanozyme are used to design signal probes by loading duplex-like DNA scaffold. Owing to the specific recognition of Ag+ toward cytosine (C) base-rich DNA, PorMOF at the distal end was close to the underlying electrode via C-Ag+-C formation, leading to an enhanced current of catalytic hydroxylamine oxidation for signal generation. Using the positive correlation between current response and Ag+ level, the electrochemical system provides a promising means for on-line monitoring of Ag+ in Lentinus edodes with recoveries from 92.8% to 106.4% and relative standard deviation from 3.98% to 8.24%, verifying the applicability of the electrochemical aptasensor toward Ag+ in Lentinus edodes. Significance and noveltyWith merits of portability, simple operation, and rapid response, the electrochemical pattern offers a useful solution for on-line monitoring of Ag+ in Lentinus edodes. By altering the DNA sequence, the proposed aptasensor provides a powerful way for monitoring other heavy metals, capable of protecting medicament production from heavy metal pollution.

The evolution of nitrogen transformation microorganism consortium under continued manganese domestication conditions

Manganese redox-stimulated bioremediation of nitrogen wastewater is receiving increasing attention. However, the nitrogen metabolic capacity and community evolution during manganese-mediated nitrogen transformation process under continued manganese domestication conditions are ambiguous. In this study, nitrogen- metabolizing microbial consortiums were incubated with synthesized Mn-humic acid complex (Mn-HA) for one month (M1), three months (M2) and six months (M3), respectively. During the Mn-HA incubation period, Bio-MnOx accompanying with bacterial consortiums (MnOB consortiums) with high TIN removal capacities were obtained. The TIN removal rates in M1, M2 and M3 were 0.220, 1.246 and 4.237 mg·L−1·h−1, respectively, which were 15.961, 90.006 and 1550.006 times higher than CK (Control Check group, no Mn-HA added group) (0.014 mg·L−1·h−1), respectively. Functional genes (amoA, AMX and narG) were most abundant in M3, which was associated with the highest nitrogen removal rate in M3. MnOB1 (bacterial consortium in M1), including Geobactor, Geothrix, Anaeromyxobacter and Bacillus, may be responsible for the Mnammox-NDMO (MnOx reduction coupled to ammonium oxidation – nitrate/nitrite-dependent low-valent Mn oxidation) process. MnOB3 (bacterial consortium in M2) enriched nitrifying bacteria Ellin6067, and denitrifying bacteria Denitratisoma, which dominated nitrogen transformation. MnOB6 (bacterial consortium in M3) enriched denitrifiers Denitratisoma, nitrifiers Ellin6067 and potential anammox bacteria SM1A02, Candidatus_Brocadia. Combined with the reduced abundance of Nitrospirae, a short-cut partial nitrification and denitrification (PND) or partial nitrification, denitrification and anammox (PNDA) could occurred in M2 and M3. It is suggested that community may have evolved into an energetically efficient short-cut nitrification, denitrification and anammox consortium to replace the full-range nitrification and denitrification community in M1 and CK under the continued manganese domestication conditions. Enhanced metabolic pathways of hydroxylamine oxidation and the nitric oxide reduction may confirm that PND or PNDA occurred in M2 and M3.

Role of Nitric Oxide in Hydroxylamine Oxidation by Ammonia-Oxidizing Bacteria.

An important role of nitric oxide (NO) as either a free intermediate in the NH3 oxidation pathway or a potential oxidant for NH3 or NH2OH has been proposed for ammonia-oxidizing bacteria (AOB) and archaea (AOA), respectively. However, tracing NO metabolism at low concentrations remains notoriously difficult. Here, we use electrochemical sensors and the mild NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO) to trace apparent NO concentration and determine production rates at low micromolar concentrations in the model AOB strain Nitrosomonas europaea. In agreement with previous studies, we found that PTIO does not affect NH3 oxidation instantaneously in both Nitrosospira briensis and Nitrosomonas europaea, unlike inhibitors for ammonia oxidation such as allylthiourea and acetylene, although it effectively scavenged NO from the cell suspensions. Quantitative analysis showed that NO production by N. europaea amounted to 3.15% to 6.23% of NO2- production, whereas N. europaea grown under O2 limitation produced NO equivalent to up to 40% of NO2- production at high substrate concentrations. In addition, we found that PTIO addition to N. europaea grown under O2 limitation abolished N2O production. These results indicate different turnover rates of NO during NH3 oxidation under O2-replete and O2-limited growth conditions in AOB. The results suggest that NO may not be a free intermediate or remain tightly bound to iron centers of enzymes during hydroxylamine oxidation and that only NH3 saturation and adaptation to O2 limitation may lead to significant dissociation of NO from hydroxylamine dehydrogenase. IMPORTANCE Ammonia oxidation by chemolithoautotrophic ammonia-oxidizing bacteria (AOB) is thought to contribute significantly to global nitrous oxide (N2O) emissions and leaching of oxidized nitrogen, particularly through their activity in nitrogen (N)-fertilized agricultural production systems. Although substantial efforts have been made to characterize the N metabolism in AOB, recent findings suggest that nitric oxide (NO) may play an important mechanistic role as a free intermediate of hydroxylamine oxidation in AOB, further implying that besides hydroxylamine dehydrogenase (HAO), additional enzymes may be required to complete the ammonia oxidation pathway. However, the NO spin trap PTIO was found to not inhibit ammonia oxidation in AOB. This study provides a combination of physiological and spectroscopic evidence that PTIO indeed scavenges only free NO in AOB and that significant amounts of free NO are produced only during incomplete hydroxylamine oxidation or nitrifier denitrification under O2-limited growth conditions.

Enhancing nitrogen removal efficiency and anammox metabolism in microbial electrolysis cell coupled anammox through different voltage application

The slow growth and difficulty in cultivating anammox bacteria limit the rapid start-up of anammox process and effective microbial enrichment. In this study, microbial electrolysis cell (MEC) was coupled with anammox to investigate the effects of different applying voltage methods on substrate removal efficiency and rates, microbial community structure, anammox metabolism and metabolic pathways. The results showed that applying voltage not only improved NH4+-N removal efficiency and removal rates, but also promoted electron transfer efficiency, key enzyme activity and extracellular polymeric substances (EPS) secretion in the systems. Step-up voltage was more conducive to the growth of Candidatus_Kuenenia in the cathode, which promoted the rapid start-up of anammox and treating wastewater with low ammonia concentration. The main metabolic pathway in step-up voltage operation was hydrazine to nitrogen, while in constant voltage operation was hydroxylamine oxidation pathway. These findings provided a new insight into the enhancement and operation of anammox system.

Pathways of N2O production by marine ammonia-oxidizing archaea determined from dual-isotope labeling

The ocean is a net source of the greenhouse gas and ozone-depleting substance, nitrous oxide (N2O), to the atmosphere. Most of that N2O is produced as a trace side product during ammonia oxidation, primarily by ammonia-oxidizing archaea (AOA), which numerically dominate the ammonia-oxidizing community in most marine environments. The pathways to N2O production and their kinetics, however, are not completely understood. Here, we use 15N and 18O isotopes to determine the kinetics of N2O production and trace the source of nitrogen (N) and oxygen (O) atoms in N2O produced by a model marine AOA species, Nitrosopumilus maritimus. We find that during ammonia oxidation, the apparent half saturation constants of nitrite and N2O production are comparable, suggesting that both processes are enzymatically controlled and tightly coupled at low ammonia concentrations. The constituent atoms in N2O are derived from ammonia, nitrite, O2, and H2O via multiple pathways. Ammonia is the primary source of N atoms in N2O, but its contribution varies with ammonia to nitrite ratio. The ratio of 45N2O to 46N2O (i.e., single or double labeled N) varies with substrate ratio, leading to widely varying isotopic signatures in the N2O pool. O2 is the primary source for O atoms. In addition to the previously demonstrated hybrid formation pathway, we found a substantial contribution by hydroxylamine oxidation, while nitrite reduction is an insignificant source of N2O. Our study highlights the power of dual 15N-18O isotope labeling to disentangle N2O production pathways in microbes, with implications for interpretation of pathways and regulation of marine N2O sources.

The contribution shift of ammonia-oxidizing archaea and bacteria to ammoxidation under Ag-NPs/SWCNTs/PS-NPs stressors in constructed wetlands

Ammonia-oxidizing microorganisms (AOMs, archaea (AOA) and bacteria (AOB)) are primarily responsible for the ammoxidation in constructed wetlands (CWs). However, little is known about evaluating the response of AOA and AOB to engineered nanomaterials (ENMs) and quantifying the shift of their contribution to ammoxidation. Herein, we operated a series of CWs exposing to silver nanoparticles (Ag-NPs), single-walled carbon nanotubes (SWCNTs), and polystyrene nano-sized plastics (PS-NPs) with the wastewater-accumulating concentration of ENMs for 180 days. The results showed that the abundance of AOA amoA gene in situ was far lower than that of AOB, while the abundance ratio of AOA to AOB increased by 15 folds after 180-day experiment. Using DNA stable isotope probing (DNA-SIP) experiment, we found that the active AOB microbiota varied substantially but the AOA was more stable across different groups. Furthermore, the co-occurrence analysis proved that ENMs stress increased the negative coexistence pattern of AOA and AOB; predictive functional profiling showed that the ENMs enhanced the functional advantage of AOA by inhibiting AOB (mainly hydroxylamine oxidation process). Finally, the contribution of AOA increased under exposing to SWCNTs (18.35%), PS-NPs (24.92%), and Ag-NPs (32.14%) compared with control group (0.03%) for 180 days. Despite this, AOB was still the primary executant of ammoxidation in CWs. Overall, in our study, the differences in activities and contributions of AOMs were quantified in CWs, and a significantly negative coexistence relationship between AOA and AOB was revealed when exposed to emerging nanomaterials.

Dynamic emissions of N<sub>2</sub>O from solid waste landfills: A review

Nitrous oxide (N<sub>2</sub>O) is an important greenhouse gas (GHG) and an ozone-depleting substance that can be emitted from landfills. Understanding the dynamics of N<sub>2</sub>O and its contribution to total emissions is critical to effective mitigation measures. This study outlines the dynamics of N<sub>2</sub>O in solid waste landfills and N<sub>2</sub>O emissions from them. N<sub>2</sub>O generation in anaerobic landfills is primarily due to denitrification and, to a lesser extent, nitrification, which occurs in the oxygenated cover soil layer and the working face. However, nitrification and denitrification are very limited within landfills. The landfill leachate contains high concentrations of ammonia (NH<sub>3</sub>). Thus, a significant amount of N<sub>2</sub>O is generated from aerobic processes during leachate treatment. Bioreactor landfills emit more N<sub>2</sub>O than traditional anaerobic landfills. The majority of N<sub>2</sub>O emitted from bioreactor landfills is generated through different pathways, such as hydroxylamine (NH<sub>2</sub>OH) oxidation, nitrifier denitrification, and heterotrophic denitrification. These processes are affected by several factors, including the carbon-to-nitrogen (C/N) ratio, NH<sub>3</sub> oxidation rate NH<sub>3</sub> oxidation rate, redox conditions, and temperature. Addressing N<sub>2</sub>O emissions from landfills will be necessary to achieve an integrated nitrogen management strategy that helps minimize N<sub>2</sub>O emissions.

Direct ammonium oxidation to nitrogen gas (Dirammox) in Alcaligenes strain HO-1: The electrode role

It has been recently suggested that Alcaligenes use a previously unknown pathway to convert ammonium into dinitrogen gas (Dirammox) via hydroxylamine (NH2OH). This fact alone already implies a significant decrease in the aeration requirements for the process, but the process would still be dependent on external aeration. This work studied the potential use of a polarised electrode as an electron acceptor for ammonium oxidation using the recently described Alcaligenes strain HO-1 as a model heterotrophic nitrifier. Results indicated that Alcaligenes strain HO-1 requires aeration for metabolism, a requirement that cannot be replaced for a polarised electrode alone. However, concomitant elimination of succinate and ammonium was observed when operating a previously grown Alcaligenes strain HO-1 culture in the presence of a polarised electrode and without aeration. The usage of a polarised electrode together with aeration did not increase the succinate nor the nitrogen removal rates observed with aeration alone. However, current density generation was observed along a feeding batch test representing an electron share of 3% of the ammonium removed in the presence of aeration and 16% without aeration. Additional tests suggested that hydroxylamine oxidation to dinitrogen gas could have a relevant role in the electron discharge onto the anode. Therefore, the presence of a polarised electrode supported the metabolic functions of Alcaligenes strain HO-1 on the simultaneous oxidation of succinate and ammonium.

Alcohols as inhibitors of ammonia oxidizing archaea and bacteria.

Ammonia oxidizers are key players in the global nitrogen cycle and are responsible for the oxidation of ammonia to nitrite, which is further oxidized to nitrate by other microorganisms. Their activity can lead to adverse effects on some human-impacted environments, including water pollution through leaching of nitrate and emissions of the greenhouse gas nitrous oxide (N2O). Ammonia monooxygenase (AMO) is the key enzyme in microbial ammonia oxidation and shared by all groups of aerobic ammonia oxidizers. The AMO has not been purified in an active form, and much of what is known about its potential structure and function comes from studies on its interactions with inhibitors. The archaeal AMO is less well studied as ammonia oxidizing archaea were discovered much more recently than their bacterial counterparts. The inhibition of ammonia oxidation by aliphatic alcohols (C1-C8) using the model terrestrial ammonia oxidizing archaeon 'Candidatus Nitrosocosmicus franklandus' C13 and the ammonia oxidizing bacterium Nitrosomonas europaea was examined in order to expand knowledge about the range of inhibitors of ammonia oxidizers. Methanol was the most potent specific inhibitor of the AMO in both ammonia oxidizers, with half-maximal inhibitory concentrations (IC50) of 0.19 and 0.31 mM, respectively. The inhibition was AMO-specific in 'Ca. N. franklandus' C13 in the presence of C1-C2 alcohols, and in N. europaea in the presence of C1-C3 alcohols. Higher chain-length alcohols caused non-specific inhibition and also inhibited hydroxylamine oxidation. Ethanol was tolerated by 'Ca. N. franklandus' C13 at a higher threshold concentration than other chain-length alcohols, with 80 mM ethanol being required for complete inhibition of ammonia oxidation.

Outer coordination sphere influences on cofactor maturation and substrate oxidation by cytochrome P460.

Product selectivity of ammonia oxidation by ammonia-oxidizing bacteria (AOB) is tightly controlled by metalloenzymes. Hydroxylamine oxidoreductase (HAO) is responsible for the oxidation of hydroxylamine (NH2OH) to nitric oxide (NO). The non-metabolic enzyme cytochrome (cyt) P460 also oxidizes NH2OH, but instead produces nitrous oxide (N2O). While both enzymes use a heme P460 cofactor, they selectively oxidize NH2OH to different products. Previously reported structures of Nitrosomonas sp. AL212 cyt P460 show that a capping phenylalanine residue rotates upon ligand binding, suggesting that this Phe may influence substrate and/or product binding. Here, we show via substitutions of the capping Phe in Nitrosomonas europaea cyt P460 that the bulky phenyl side-chain promotes the heme-lysine cross-link forming reaction operative in maturing the cofactor. Additionally, the Phe side-chain plays an important role in modulating product selectivity between N2O and NO during NH2OH oxidation under aerobic conditions. A picture emerges where the sterics and electrostatics of the side-chain in this capping position control the kinetics of N2O formation and NO binding affinity. This demonstrates how the outer coordination sphere of cyt P460 is tuned not only for selective NH2OH oxidation, but also for the autocatalytic cross-link forming reaction that imbues activity to an otherwise inactive protein.

Natural dried Vanilla Beans powder (VBP) mediated CuO nanoparticles catalysis: Aerobic oxidation of hydroxylamines from industrial waste water using ecofriendly method

In an industrial waste water treatment plant, a novel efficient approach is employed to achieve quantitation of hydroxylamines. For aerobic alcohol oxidation, Vanilla Beans Powder (VBP) mediated Copper oxide nanoparticles were created. Over the CuO NPs, various Hydroxylamines alcohol substrates were converted to the appropriate aldehyde in excellent yield. XRD, XPR, FTIR, UV, TEM, EDX, TGA, and DTA were all used to characterise CuO NPs. Metal nanoparticle stabilisation and oxidation state management are crucial features of nanocatalysis. The CuO NPs was validated by XRD peaks are linked to copper's cubic crystalline structures (002), (111), (202), (202), and (113), and the HR TEM research 20 nm image of a small segment displayed clear lattice fringe separations between 0.250 nm and 0.226 nm, which correspond to CuO (002)'s fcc (face-centered cubic). The XPS analysis of Cu O NPs survey scan of CuO-Nps revealed the presence of Cu, O, and C without any other impurity in the nanoparticles, Cu 2p3/2 and Cu 2p1/2, with binding energies of 931.26 and 950.97 eV and a spin-orbit splitting of 19.71 eV, respectively, were identified in the XP spectra of the Cu 2p electrons. Catalytic activity of CuONPs were used various concentration for conversion of hydroxylamines, for example 2-((2-amino ethyl) amino)ethanol (1a) is converted to 2-((2-aminoethyl)amino)acetaldehyde. CuO NPs (6%) of catalyst was performed well, it is 98% recovery and 100% conversation of product with 4 mints compared with Vanilla Beans powder (VBP) and vaniline. In addition to (FT-IR, 1 H, 13 C NMR, GC-MS) and elemental analysis was performed on the isolated product. The new volatile compounds was analyzed by GC-MS and also fragmentation studies discussed in this work. These stabilization and frequentation were well described. • Vanilla Beans powder mediated Cupper oxide nanoparticles were used to obtain heterogeneous catalysts. • Removal of Industrial wastewaters contamination of β -Hydroxylamines • Efficient procedure for the Aerobic oxidation of oxidation of β -Hydroxylamines in the presence of CuO NPs catalyst. • Efficient recyclability of the catalyst.

Diversity of the Hydroxylamine Oxidoreductase (HAO) Gene and Its Enzyme Active Site in Agricultural Field Soils.

Nitrification is a key process in the biogeochemical nitrogen cycle and a major emission source of the greenhouse gas nitrous oxide (N2O). The periplasmic enzyme hydroxylamine oxidoreductase (HAO) is involved in the oxidation of hydroxylamine to nitric oxide in the second step of nitrification, producing N2O as a byproduct. Its three-dimensional structure demonstrates that slight differences in HAO active site residues have inhibitor effects. Therefore, a more detailed understanding of the diversity of HAO active site residues in soil microorganisms is important for the development of novel nitrification inhibitors using structure-guided drug design. However, this has not yet been examined. In the present study, we investigated hao gene diversity in beta-proteobacterial ammonia-oxidizing bacteria (β-AOB) and complete ammonia-oxidizing (comammox; Nitrospira spp.) bacteria in agricultural fields using a clone library ana-lysis. A total of 1,949 hao gene sequences revealed that hao gene diversity in β-AOB and comammox bacteria was affected by the fertilizer treatment and field type, respectively. Moreover, hao sequences showed the almost complete conservation of the six HAO active site residues in both β-AOB and comammox bacteria. The diversity of nitrifying bacteria showed similarity between hao and amoA genes. The nxrB amplicon sequence revealed the dominance of Nitrospira cluster II in tea field soils. The present study is the first to reveal hao gene diversity in agricultural soils, which will accelerate the efficient screening of HAO inhibitors and evaluations of their suppressive effects on nitrification in agricultural soils.

Effects of hydroxylamine on treatment of anaerobic digestate of pig manure in partial nitrification-anaerobic ammonium oxidation

Partial nitrification-anaerobic ammonium oxidation (PN-anammox) was started up within 40 days by bioaugmentation and aeration control, and its performance in the treatment of anaerobic digestate of pig manure (ADPM) was evaluated. Inhibitors in ADPM decreased the nitrogen removal rate (NRR) by 0.24 g N/L/d. The effect and mechanism of hydroxylamine (NH2OH) alleviation of PN-anammox inhibition during ADPM treatment were investigated. As an intermediate product of anammox and ammonia-oxidizing bacteria, NH2OH strengthened energy metabolism, improved the activity and abundance of functional bacteria, and eliminated miscellaneous bacteria, increasing the average NRR by 31%. However, the average nitrous oxide emission was increased by 10.1% via hydroxylamine oxidation. The results showed that synergy and competition among nitrogen-transforming microorganisms were crucial for NRR and that NH2OH played an essential role in maintaining efficient operation. This study lays a foundation for restoring PN-anammox for treating livestock wastewater.