Laccase-induced decontamination and humification mechanisms of estrogen in water-crop matrices.

Enzymatic humification plays a crucial biogeochemical role in eliminating steroidal estrogens and expanding organic carbon stocks. Estrogenic contaminants in agroecosystems can be taken up and acropetally translocated by crops, but the roles of laccase-triggered rhizospheric humification (L-TRH) in pollutant dissipation and plant uptake remain poorly understood. In this study, the laccase-induced decontamination and humification mechanisms of 17ß-estradiol (E2) in water-crop media were investigated by performing greenhouse pot experiments with maize seedlings (Zea mays L.). The results demonstrated that L-TRH effectively dissipated E2 in the rhizosphere solution and achieved the kinetic constants of E2 dissipation at 10 and 50 µM by 8.05 and 2.75 times as much as the treatments without laccase addition, respectively. The copolymerization of E2 and root exudates (i.e. phenols and amino acids) consolidated by L-TRH produced a larger amount of humified precipitates with the richly functional carbon architectures. The growth parameters and photosynthetic pigment levels of maize seedlings were greatly impeded after a 120-h exposure to 50 µM E2, but L-TRH motivated the detoxication process and thus mitigated the phytotoxicity and bioavailability of E2. The tested E2 contents in the maize tissues initially increased sharply with the cultivation time but decreased steadily. Compared with the treatment without laccase addition, the uptake and accumulation of E2 in the maize tissues were obviously diminished by L-TRH. E2 oligomers such as dimer, trimer, and tetramer recognized in the rhizosphere solution were also detected in the root tissues but not in the shoots, demonstrating that the acropetal translocation of E2 oligomers was interrupted. These results highlight a promising strategy for decontaminating estrogenic pollutants, boosting rhizospheric humification, and realizing low-carbon emissions, which would be beneficial for agroenvironmental bioremediation and sustainability.

Biogenic calcium improved Cd2+ and Pb2+ immobilization in soil using the ureolytic bacteria Bacillus pasteurii.

Bioremediation based on microbial-induced carbonate precipitation (MICP) was conducted in cadmium and lead contaminated soil to investigate the effects of MICP on Cd and Pb in soil. In this study, soil indigenous nitrogen was shown to induce MICP to stabilize heavy metals without inputting exogenous urea. The results showed that applying Bacillus pasteurii coupled with CaCl2 reduced Cd and Pb bioavailability, which could be clarified through the proportion of exchangeable Cd and Pb in soil decreasing by 23.65 % and 12.76 %, respectively. Moreover, B. pasteurii was combined separately with hydroxyapatite (HAP), eggshells (ES), and oyster shells (OS) to investigate their effects on soil heavy metals' chemical fractions, toxicity characteristic leaching procedure (TCLP)-extractable Cd and Pb as well as enzymatic activity. Results showed that applying B. pasteurii in soil significantly decreased the heavy metals in the exchangeable fraction and increased them in the carbonate phase fraction. When B. pasteurii was combined with ES and OS, the content of carbonate-bound Cd increased by 114.72 % and 118.81 %, respectively, significantly higher than when B. pasteurii was combined with HAP, wherein the fraction of carbonate-bound Cd increased by 86 %. The combination of B. pasteurii and biogenic calcium effectively reduced the leached contents of Cd and Pb in soil, and the TCLP-extractable Cd and Pb fractions decreased by 43.88 % and 30.66 %, respectively, in the BP + ES group and by 52.60 % and 41.77 %, respectively, in the BP + OS group. This proved that MICP reduced heavy metal bioavailability in the soil. Meanwhile, applying B. pasteurii and calcium materials significantly increased the soil urease enzyme activity. The microstructure and chemical composition of the soil samples were studied, and the results from scanning electron microscope, Fourier transform infra-red spectroscopy, and X-ray diffraction demonstrated the MICP process and identified the formation of CaCO3, Ca0.67Cd0.33CO3, and PbCO3 in heavy metal-contaminated soil.

Enhanced microbial degradation mediated by pyrogenic carbon toward p-nitrophenol: Role of carbon structures and iron minerals.

Pyrogenic carbon (PC) including black carbons and engineered carbons can mediate the extracellular electron transfer to facilitate the biogeochemical reaction with organic pollutants. Yet, the role of carbon structures and iron minerals on PC-mediated microbial degradation is still lacking of understanding. Herein, we studied the electrochemical properties of PCs produced from varied feedstock with regards to the mediated degradation of p-nitrophenol (PNP) by Shewanella putrefaciens CN32 in anoxic system. Mediated degradation by PCs was enhanced by facilitating extracellular electron transfer through oxygenated group and graphitic structure. Graphitic crystallites improved the electron-accepting capacity (as suggested by ID/IG and EAC) and diminished the electrochemical impedance (as suggested by Rct), contributing to PNP degradation under the anoxic system. Furthermore, more interfacial adsorption was conducive to the mediated reduction by the graphitic structure on PCs of high-temperature. In the presence of iron minerals, both hematite and goethite significantly facilitated PC-mediated degradation, which could be ascribed to the enhancement of the electron-donating capacity of microorganism and the accumulation of the reductive-state PCs by the interaction with generated Fe(II). This work paves a feasible way to the technical design on the remediation of phenolic contaminants by PC-mediated microbial degradation in environment.

Enhanced generation of reactive oxygen species by pyrite for As(III) oxidation and immobilization: The vital role of Fe(II).

The migration and conversion of arsenic in the environment usually accompany by the redox of iron-bearing minerals. For instance, the oxidation of pyrite can generate reactive oxygen species (ROS) affecting the species of arsenic, but the types and roles of ROS have been unclear. This paper demonstrated the vital role of Fe(II) in the pyrite for the formation of ROS. Results showed that exogenous addition of Fe(II) significantly enhanced the removal rate of As(III) by pyrite. 2,2'-bipyridine (BPY) decreased the oxidation of As(III) by complexing with Fe2+ in solution, whilst EDTA enhanced the oxidation of As(III) by boosting the autoxidation of Fe2+. In addition, neutral pH is superior for the oxidation of As(III) and removal of total arsenic. Importantly, Methanol, SOD enzyme and PMOS inhibited 54%, 28% and 17.5% of As(III) oxidation, respectively, which indicated O2•- and •OH were the main contributors to As(III) oxidation, and Fe(IV) contributed a small part of As(III) oxidation. The content of As(V) in the FeS2-Fe2+-As(III) system was higher than that in the FeS2-As(III) system, further confirming the vital role of Fe(II) for As(III) oxidation. Lepidocrocite was produced in a single Fe2+ system, which was not detected in the FeS2-As(III) system. Thus, the presence of mineral surfaces changed the oxidation products of Fe2+ and accelerated the oxidation and immobilization of As(III). FA (Fulvic Acid) and HA (Humic Acid) accelerated the oxidation of As(III), but the oxidation of As(III) by pyrite was inhibited to a certain extent, with increasing phenolic hydroxyl groups in phenolic acid. Our findings provide new insight into the oxidative species in the pyrite-Fe(II) system and will help guide the remediation of arsenic pollution in complex environmental systems.

Exolaccase-boosted humification for agricultural applications.

Globally, phenolic contaminants have posed a considerable threat to agro-ecosystems. Exolaccase-boosted humification may be an admirable strategy for phenolic detoxification by creating multifunctional humic-like products (H-LPs). Nonetheless, the potential applicability of the formed H-LPs in agricultural production is still overlooked. This review describes immobilized exolaccase-enabled humification in eliminating phenolic pollutants and producing artificial H-LPs. The similarities and differences between artificial H-LPs and natural humic substances (HSs) in chemical properties are compared. In particular, the agronomic effects of these reproducible artificial H-LPs are highlighted. On the basis of the above summary, the granulation process is employed to prepare granular humic-like organic fertilizers, which can be applied to field crops by mechanical side-deep fertilization. Finally, the challenges and perspectives of exolaccase-boosted humification for practical applications are also discussed. This review is a first step toward a more profound understanding of phenolic detoxification, soil improvement, and agricultural production by exolaccase-boosted humification.

Inhibition mechanisms of Fe2+/Fe3+ and Mn2+ on fungal laccase-enabled bisphenol a polyreaction.

Bisphenol A (BPA) is regarded as an endocrine disruptor associated with negative health effects in animals and humans. Laccase from white-rot fungus can enable BPA oxidation and auto-polymerization to circumvent its biotoxicity, but the work concerning the effect mechanisms of divalent and trivalent metal ions (MIs) on BPA polyreaction have rarely been reported. Herein, Trametes versicolor laccase-started BPA conversion within 1 h followed pseudo-first order kinetics, and the rate constant (kprcs) and half-life were respectively 0.61 h-1 and 1.14 h. The presence of Ca2+, Mg2+, Cu2+, Pb2+, Cd2+, Zn2+ and Al3+ exhibited insignificant impact on BPA removal, whereas Fe2+, Fe3+ and Mn2+ had a strong inhibiting effect. Compared with MI-free, the kprcs values of BPA respectively lowered 34.4%, 44.3% and 98.4% in the presence of Fe2+, Fe3+ and Mn2+. Enzymatic activity and differential absorption spectrum disclosed that the inhibitory actions were accomplished by two different mechanisms. One is Fe2+ was preferentially oxidized into Fe3+ that restrained laccase activity at the initial stage of reaction, and subsequently, the formed Fe3+ complex bound with laccase T1-Cu site and thus impeded the single-electron transfer system. The other is Mn2+ was instantly oxidized by laccase to generate Mn3+-citrate complex, which completely consumed the dissolved O2 in solution and consequently terminated BPA removal. Considering environmental bioremediation, T. versicolor laccase-enabled auto-polymerization is a simple and convenient candidate to eliminate BPA in enzymatic wastewater treatment, however the effects of Fe2+/Fe3+ and Mn2+ on BPA decontamination should be cautiously assessed.

Arsenic and cadmium bioavailability to rice (Oryza sativa L.) plant in paddy soil: Influence of sulfate application.

Arsenic (As) and cadmium (Cd) accumulate easily in rice grains that pose a non-negligible threat to human health worldwide. Sulfur fertilizer has been shown to affect the mobilization of As and Cd in paddy soil, but the effect of co-contamination by As and Cd has not been explored. This study selected three soils co-contaminated with As and Cd from Shangyu (SY), Tongling (TL) and Ma'anshan (MA). Incubation experiments and pot experiments were carried out to explore the effect of sulfate supply (100 mg kg-1) on the bioavailability of As and Cd in soil and the rice growth. The results showed that the exogenous sulfate decreased As concentrations in porewater of SY and TL by 51.1% and 29.2% through forming arsenic-sulfide minerals. The exchangeable Cd in soil also declined by 25.6% and 18.6% and transformed into Fe and Mn oxides-bound Cd. The relative abundance of Desulfotomaculum, Desulfurispora and dsr gene increased remarkably indicated that sulfate addition stimulated the activity of sulfate-reducing bacteria. In MA soil, sulfate addition immobilized Cd but had little effect on As solubility, which was speculated to be related to the high sulfate background of the soil. Further pot experiments showed that sulfate application significantly increased rice tillers, biomass, chlorophyll content in shoots, and decreased electrolyte leakage in root. Finally, sulfate significantly reduced As and Cd in SY rice shoots by 60.2% and 40.8%, respectively, while As decreased by 39.6% in TL rice shoots and Cd decreased by 23.0% in MA rice shoots. These results indicate that the application of sulfate can reduce the bioavailability of As and Cd in the soil-rice system and promote rice growth, and it is possible to reduce the accumulation of As and Cd in rice plants simultaneously.

New insights into humic acid-boosted conversion of bisphenol A by laccase-activated co-polyreaction: Kinetics, products, and phytotoxicity.

How humic acid (HA) modifies bisphenol A (BPA) conversion in exoenzyme-activated polyreaction is poorly understood. Herein, the influencing mechanism of HA on laccase-induced BPA self-polymerization was investigated, and the phytotoxicity of the produced BPA self/co-polymers was assessed for the first time. HA prominently boosted BPA elimination, and the rate constants of BPA conversion augmented from 0.61 to 1.43 h-1 as HA level raised from 0 to 50 mg·L-1. It is because the generated BPA-HA co-polymers promptly lowered the yields of long-chain BPA self-oligomers, consequently maintaining laccase activity through opening enzymatic substrate-binding pockets. Notably, a few BPA monomers were re-released from the loosely bound self-polymers and co-polymers, and the releasing amounts respectively were 13.9 - 22.4% and 0.3 - 0.5% at pH 2 - 11. Formation of self/co-polymers was greatly conducive to avoiding BPA biotoxicity. Compared with BPA self-polymers, the phytotoxicity of BPA co-polymers to germinated radish (Raphanus sativus L.) seeds was lower due to these covalently bound products were more complex and stable. It follows that laccase-mediated co-polymerization played a significant role in BPA conversion, contaminant detoxification, and carbon sequestration. These findings are not only beneficial to clarifying exoenzyme-activated the generation mechanism of BPA co-polymers in water, but to reusing these supramolecular aggregates in crop growth.

Insights into the Applications of Extracellular Laccase-Aided Humification in Livestock Manure Composting.

Traditional composting is a well-suited biotechnology for on-farm management of livestock manure (LM) but still leads to the release of toxic micropollutants and imbalance of nutrients. One in situ exoenzyme-assisted composting has shown promise to ameliorate the agronomical quality of end products by improving humification and polymerization. The naturally occurring extracellular laccase from microorganisms belongs to a multicopper phenoloxidase, which is verified for its versatility to tackle micropollutants and conserve organics through the reactive radical-enabled decomposition and polymerization channels. Laccase possesses an indispensable relationship with humus formation during LM composting, but its potential applications for the harmless disposal and resource utilization of LM have until now been overlooked. Herein, we review the extracellular laccase-aided humification mechanism and its optimizing strategy to maintain enzyme activity and in situ production, highlighting the critical roles of laccase in treating micropollutants and preserving organics during LM composting. Particularly, the functional effects of the formed humification products by laccase-amended composting on plant growth are also discussed. Finally, the future perspectives and outstanding questions are summarized. This critical review provides fundamental insights into laccase-boosted humification that ameliorates the quality of end products in LM composting, which is beneficial to guide and advance the practical applications of exoenzyme in humification remediation, the carbon cycle, and agriculture protection.

Foliar application of anthocyanin extract regulates cadmium accumulation and distribution in rice (Oryza sativa L.) at tillering and booting stages.

Anthocyanin extract has been applied in agricultural production and enhanced tolerance of plants to adverse effects of Cd stress. Rice was subjected to different concentration of Cd and blueberry anthocyanin, and the effects on rice growth, antioxidative defense, Cd distribution in rice tissues, FTIR and TEM characterization of rice leaves were examined to explain the Cd reductions in rice grains and the protective mechanisms by blueberry anthocyanin. Foliar spray of blueberry anthocyanin at tillering and booting stages was effective for reducing Cd concentration in rice grains and increasing the rice yield, anthocyanin and Cd concentration of rice leaves under 1.0 and 10.0 mg/kg Cd stress. The Cd concentration in rice grains was less than the China national standard of Cd for rice grains (0.2 mg/kg) after surface spraying by 5.0-12.5 g/L blueberry anthocyanin under 1.0 mg/kg Cd stress, while 7.5 g/L spray concentration was the best choice. Blueberry anthocyanin spraying prevents Cd from being transferred from leaves to rice grains mainly by fixation of Cd in soluble and organelle fractions at the tillering and booting stages, and reduces H2O2 and MDA accumulation in rice leaves to decrease Cd toxicity. Combined with FTIR and TEM characterization of rice leaves, the results indicated that surface spraying of 7.5 g/L blueberry anthocyanin under 1.0 mg/kg Cd stress could effectively relieve Cd oxidative damage, and form chelates with Cd ions to immobilize Cd in rice leaves. Hence, blueberry anthocyanin could be used as a foliar resistance control agent to reduce Cd accumulation in rice grains through chelate compound synthesis and decrease Cd toxicity by preventing membrane lipid peroxidation and H2O2 accumulation.

Advances in laccase-triggered anabolism for biotechnology applications.

This is the first comprehensive overview of laccase-triggered anabolism from fundamental theory to biotechnology applications. Laccase is a typical biological oxidordeuctase that induces the one-electronic transfer of diverse substrates for engendering four phenoxy radicals with concomitant reduction of O2 into 2H2O. In vivo, laccase can participate in anabolic processes to create multifarious functional biopolymers such as fungal pigments, plant lignins, and insect cuticles, using mono/polyphenols and their derivatives as enzymatic substrates, and is thus conducive to biological tissue morphogenesis and global carbon storage. Exhilaratingly, fungal laccase has high redox potential (E° = 500-800 mV) and thermodynamic efficiency, making it a remarkable candidate for utilization as a versatile catalyst in the green and circular economy. This review elaborates the anabolic mechanisms of laccase in initiating the polymerization of natural phenolic compounds and their derivatives in vivo via radical-based self/cross-coupling. Information is also presented on laccase immobilization engineering that expands the practical application ranges of laccase in biotechnology by improving the enzymatic catalytic activity, stability, and reuse rate. Particularly, advances in biotechnology applications in vitro through fungal laccase-triggered macromolecular biosynthesis may provide a key research direction beneficial to the rational design of green chemistry.

Fungal laccase-triggered 17ß-estradiol humification kinetics and mechanisms in the presence of humic precursors.

Naturally-occurring phenolic acids (PAs) act as humic precursors that participate in the conversion behaviors and coupling pathways of steroidal estrogens (SEs) during laccase-triggered humification processes (L-THPs). Herein, the influences and mechanisms of PAs on Trametes versicolor laccase-evoked 17ß-estradiol (E2) conversion kinetics and humification routes were explored. Fungal laccase was fleet in converting > 99% of E2, and the calculated pseudo-first-order velocity constant and half-time values were respectively 0.039 min-1 and 17.906 min. PAs containing an O-dihydroxy moiety such as gallic acid and caffeic acid evidently hampered E2 humification owning to the yielded highly reactive O-quinones reversed E2 radicals by hydrogen transfer mechanism, implying that the inhibition effect was enormously dependent upon the number and position of the phenolic -OH present in humic precursors. Oligomers and polymers with carbon-carbon/oxygen links were tentatively found as E2 main humified species resulting from laccase-evoked successive oxidative-coupling. Note that PAs participating in the humification also encountered oxydehydrogenation, self-polymerization, and cross-binding to E2. Interestingly, the -COOH and -OCH3 groups of PAs could be deprived in radical-caused self/co-polymerization. The generation of humified products not only circumvented the environmental risks of parent compounds but accelerated global carbon sequestration. To our knowledge, this is the first in-depth revelation of the humification pathways and related mechanisms of SEs with humic precursors in aquatic ecosystems by L-THPs.

Surface spraying of anthocyanin through antioxidant defense and subcellular sequestration to decrease Cd accumulation in rice (Oryza sativa L.) grains in a lead-zinc mine area.

As an important class of flavonoids, anthocyanin has been used to enhance plant-defensive mechanisms against heavy metal stress. However, there are few available reports regarding surface spraying of anthocyanin for reduction of Cd poisoning in rice and its practical applications in paddy fields. After rice growing, measurements were taken of rice growth, pigments, the antioxidant system, thiol compounds, and distribution of Cd in rice tissues. The results showed that surface spraying anthocyanin could promote rice growth, and relative to the control, total chlorophyll significantly increased by 22.62% after surface spraying of 7.5 g L-1 anthocyanin. Simultaneously, Cd accumulation in rice grains was 0.17 ± 0.02 mg kg-1, which was significantly decreased by 46.88% relative to the control. In the pot experiment (40-day-old rice), treatment with 7.5 g L-1 anthocyanin resulted in decreases of ·O2-, H2O2, and malondialdehyde contents in rice leaves, while the activities of superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase were increased by 59.10, 23.81, 41.75, and 9.39%, respectively. Meanwhile, contents of glutathione, ascorbic acid, non-protein thiols, and phytochelatins showed respective increases of 7.24, 14.49, 42.81, and 41.13% compared with the control value. Subcellular analysis revealed that surface spraying of anthocyanin increased organelle and soluble fractions of Cd in leaf cells. In conclusion, surface spraying of 7.5 g L-1 anthocyanin was mainly attributed to increased antioxidant activities and subcellular sequestration of Cd in organelles and soluble fractions in rice leaves to reduce Cd accumulation in rice grains in the field.

Influence of humic acids on fungal laccase-initiated 17α-ethynylestradiol oligomerization: Transformation kinetics and products distribution.

Fungal laccase has aroused great concern in rapidly removing estrogens because of its ability to accelerate humification and oligomerization. Here, the effect of two humic acids (HAs) on the reaction kinetics and products distribution of 17α-ethynylestradiol (EE2) in laccase-initiated humification and coupling was systematically elucidated. Laccase from Trametes versicolor exhibited over 98.3% removal rate for EE2 at pH 5.0 within 120 min, while HAs invariably restrained EE2 transformation by competing with target-substrate for the enzymatic catalytic center. EE2 removal followed pseudo-first-order kinetics, and the rate constant was decreased markedly with increasing concentration of two HAs (0-60 mg L-1). Additionally, laccase heightened the aromaticity and humification degrees (A250 nm/A365 nm ratio) of HAs probably due to the formation of new humic polymers such as (HA)m and/or (HA)m-(EE2)n (m and n represent the number of HA and EE2 units, respectively). Three major EE2 oligomers were identified, in accordance with a mechanism involving the phenoxy radical-driven polymerization to yield a wide variety of self-coupling products. Notably, HAs diminished the extent of EE2 self-coupling but aggrandized the cross-coupling between EE2 and HAs, and the inhibition degree of EE2 self-coupling increased with the concentration of HAs. One major reason is EE2 could be covalently incorporated into humic molecules to produce (HA)m-(EE2)n cross-coupling products via radical-caused C-C, C-O-C, and/or C-O-C bonds, thereby reducing EE2 self-oligomerization. These findings highlight that HAs play a vital role in the fungal laccase-induced humification and oligomerization of EE2, which obviously alter the geochemical fate and transport of EE2 in natural aquatic ecosystems.

Remediation of heavy metals polluted environment using Fe-based nanoparticles: Mechanisms, influencing factors, and environmental implications.

Environmental pollution by heavy metals (HMs) has raised considerable attention due to their toxic impacts on plants, animals and human beings. Thus, the environmental cleanup of these toxic (HMs) is extremely urgent both from the environmental and biological point of view. To remediate HMs-polluted environment, several nanoparticles (NPs) such as metals and its oxides, carbon materials, zeolites, and bimetallic NPs have been documented. Among these, Fe-based NPs have emerged as an effective choice for remediating environmental contamination, due to infinite size, high reactivity, and adsorption properties. This review summarizes the utilization of various Fe-based NPs such as nano zero-valent iron (NZVI), modified-NZVI, supported-NZVI, doped-NZVI, and Fe oxides and hydroxides in remediating the HMs-polluted environment. It presents a comprehensive elaboration on the possible reaction mechanisms between the Fe-based NPs and heavy metals, including adsorption, oxidation/reduction, and precipitation. Subsequently, the environmental factors (e.g., pH, organic matter, and redox) affecting the reactivity of the Fe-based NPs with heavy metals are also highlighted in the current study. Research shows that Fe-based NPs can be toxic to living organisms. In this context, this review points out the environmental hazards associated with the application of Fe-based NPs and proposes future recommendations for the utilization of these NPs.

Isolation of Trametes hirsuta La-7 with high laccase-productivity and its application in metabolism of 17ß-estradiol.

Estrogens, which are extensive in the eco-environments, are a category of high-toxic emerging contaminants that induce metabolic disorders and even carcinogenic risks in wildlife and humans. Here we investigate whether fungus-secreted laccase can be used as a green catalyst to eliminate a representative estrogen, 17ß-estradiol (E2). A white-rot fungus Trametes hirsuta La-7 with high laccase-productivity, was isolated from pig manure-contaminated soil. Extracellular laccase activity expressed by strain La-7 was 65.4 U·mL-1 for a 3 d inoculation under the optimal fermentation parameters. The concentrated-crude laccase from Trametes hirsuta La-7 (CC-ThLac) was capable of effectively metabolizing E2 at pH 4-6, and the apparent pseudo first-order reaction rate constant and half-life values were respectively 0.027-0.055 min-1 and 25.86-12.67 min (R2 > 0.98). The mass measurement of high-resolution mass spectrometry in combination with 13C-isotope labeling identified that the main by-products of E2 metabolism were dimers, trimers, and tetramers, which are consistent with radical-driven C-C and/or C-O-C covalent coupling pathway, involving the initial enzymatic production of phenoxy radical intermediates and then the successive oxidative-oligomerization of radical intermediates. The formation of oligomers dramatically reduced the estrogenic activity of E2. Additionally, CC-ThLac also exhibited high-efficiency metabolism capability toward E2 in the natural water and pig manure, with more than 94.4% and 91.0% of E2 having been metabolized, respectively. These findings provide a broad prospect for the clean biotechnological applications of Trametes hirsuta La-7 in estrogen-contaminated ecosystems.

MnO2 nanozyme-driven polymerization and decomposition mechanisms of 17ß-estradiol: Influence of humic acid.

Nanozymes, which display the bifunctional properties of nanomaterials and natural enzymes, are useful tools for environmental remediation. In this research, nano-MnO2 was selected for its intrinsic enzyme-like activity to remove 17ß-estradiol (E2). Results indicated that nano-MnO2 exhibited laccase-like activity (7.22 U·mg-1) and removed 97.3 % of E2 at pH 6. Humic acid (HA) impeded E2 removal (only 72.4 %) by competing with E2 for the catalytic sites of the MnO2 nanozyme surface, and there was a good linear correlation between the kinetic constants and HA concentrations (R2 = 0.9489). Notably, the phenolic -OH of E2 interacted with HA to yield various polymeric products via radical-driven covalent coupling, resulting in ablation of phenolic -OH but increase of ether groups in the polymeric structure. Intermediate products, including estrone, E2 homo-/hetero-oligomers, E2 hydroxylated and quinone-like products, as well as aromatic ring-opening species, were identified. Interestingly, HA hindered the extent of E2 oxidation, homo-coupling, and decomposition but accelerated E2 and HA hetero-coupling. A reasonable catalytic pathway of E2 and HA involving MnO2 nanozyme was proposed. These findings provide novel insights into the influence of HA on MnO2 nanozyme-driven E2 radical polymerization and decomposition, consequently favoring the ecological water restoration and the global carbon cycle.

Silver nanoparticles inhibit denitrification by altering the viability and metabolic activity of Pseudomonas stutzeri.

The environmental toxicity of silver nanoparticles (AgNPs) is currently the focus of intensive research. However, the mechanisms underlying the cytotoxic effects of AgNPs on denitrifying microbes have yet to be explicitly demonstrated. Herein, Pseudomonas stutzeri was used to explore the effects of AgNPs on denitrification and cytotoxicity. The denitrification efficiency decreased from 94.91% in the AgNP-free treatment to 87.66%, 60.51% and 36.10% with treatments of 3.125, 6.25 and 12.5 mg/L AgNPs, respectively. The inhibition and delay in the denitrification process from treatment with AgNPs likely occurred through alteration of the viability and metabolic activity of P. stutzeri. Flow cytometry analysis indicated that the early apoptotic rates of P. stutzeri were 8.72%, 30.60%, and 48.60% with treatments of 3.125, 6.25, and 12.5 mg/L AgNPs, respectively. Results for scanning electron microscope (SEM) imaging, ζ-potential analysis, lactate dehydrogenase (LDH) release and malondialdehyde (MDA) production assays demonstrated adsorption of AgNPs on the cell surface, which altered membrane potential and mediated lipid peroxidation; these events eventually resulted in the aberration of cell morphology. Transmission electron microscopy (TEM) images and measurements of Ag content distribution by ICP-MS indicated that AgNPs were easily internalized by P. stutzeri, which increased the accumulation of reactive oxygen species (ROS). Furthermore, the presence of AgNPs also greatly inhibited expression of genes napA, nirS, cnorB, and nosZ, thereby reducing the activities of nitrate reductase (NAR) and nitrite reductase (NIR). These findings will help further our understanding of the mechanism underlying AgNPs cytotoxicity, and provide the means to evaluate the negative effect of nanoparticles in the environment.

[Effects of Silver Nanoparticles and Silver Ions on Soil Nitrification Microorganisms and Ammoxidation].

In order to study the effect of nanosilver on soil nitrification microorganisms and nitrogen transformation, soil culture experiments were carried out. Yellow brown soil and paddy soil were first spiked with different doses of nanosilver (10, 50, 100 mg·kg-1) and silver ions (1, 5, 10 mg·kg-1). Then, the number of nitrifying bacteria, activity of soil invertase, amoA gene abundance, NH4+-N content, NO3--N content, and soil potential ammonia oxidation rate were determined. The results showed that the number of nitrite bacteria and nitrate bacteria decreased significantly when the soils were treated with nanosilver and silver ions. Soil invertases were inhibited, and the effect on urease was greater than that on catalase. The amoA gene abundances of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) decreased, and the effect on the gene abundance of AOB was greater than that of AOA. When (NH4)2SO4 was added to the soil, nanosilver and silver ion pollutants caused NH4+-N to accumulate, and the contents of NO3--N were reduced, the rate of ammonia oxidation decreased, and the transformation of ammonium nitrogen to nitrate nitrogen was inhibited. This research suggests that nanosilver and silver ions can have toxic effects on soil nitrification microorganisms and ammonium nitrogen conversion, and the degree of influence was found to be related to the soil physical and chemical properties.

Oxidase-Like Catalytic Performance of Nano-MnO2 and Its Potential Application for Metal Ions Detection in Water.

Certain nano-scale metal oxides exhibiting the intrinsic enzyme-like reactivity had been used for environment monitoring. Herein, we evaluated the oxidase-mimicking activity of environmentally relevant nano-MnO2 and its sensitivity to the presence of metal ions, and particularly, the use of MnO2 nanozyme to potentially detect Cu2+, Zn2+, Mn2+, and Fe2+ in water. The results indicated the oxidase-like activity of nano-MnO2 at acidic pH-driven oxidation of 2,6-dimethoxyphenol (2,6-DMP) via a single-electron transfer process, leading to the formation of a yellow product. Notably, the presence of Cu2+ and Mn2+ heightened the oxidase-mimicking activity of nano-MnO2 at 25°C and pH 3.8, showing that Cu2+ and Mn2+ could modify the reactive sites of nano-MnO2 surface to ameliorate its catalytic activity, while the activity of MnO2 nanozyme in systems with Zn2+ and Fe2+ was impeded probably because of the strong affinity of Zn2+ and Fe2+ toward nano-MnO2 surface. Based on these effects, we designed a procedure to use MnO2 nanozyme to, respectively, detect Cu2+, Zn2+, Mn2+, and Fe2+ in the real water samples. MnO2 nanozyme-based detecting systems achieved high accuracy (relative errors: 2.2-26.1%) and recovery (93.0-124.0%) for detection of the four metal ions, respectively. Such cost-effective detecting systems may provide a potential application for quantitative determination of metal ions in real water environmental samples.