Carbon-based nanozymes: design, catalytic mechanisms, and environmental applications.

Carbon-based nanozymes are synthetic nanomaterials that are predominantly constituted of carbon-based materials, which mimic the catalytic properties of natural enzymes, boasting features such as tunable catalytic activity, robust regenerative capacity, and exceptional stability. Due to the impressive enzymatic performance similar to various enzymes such as peroxidase, superoxide dismutase, and oxidase, they are widely used for detecting and degrading pollutants in the environment. This paper presents an exhaustive review of the fundamental design principles, catalytic mechanisms, and prospective applications of carbon-based nanozymes in the environmental field. These studies not only serve to augment the comprehension on the intricate operational mechanism inherent in these synthetic nanostructures, but also provide essential guidelines and illuminating perspectives for advancing their development and practical applications. Future studies that are imperative to delve into the untapped potential of carbon-based nanozymes within the environmental domain was needed to be explored to fully harness their ability to deliver broader and more impactful environmental preservation and management outcomes.

Mechanistic Insights into the Biodegradation of Carbon Dots by Fungal Laccase.

While carbon dots (CDs) have the potential to support the agricultural revolution, it remains obscure about their environmental fate and bioavailability by plants. Fungal laccase-mediated biotransformation of carbon nanomaterials has received little attention despite its known capacity to eliminate recalcitrant contaminants. Herein, we presented the initial investigation into the transformation of CDs by fungal laccase. The degradation rates of CDs were determined to be first-order in both substrate and enzyme. Computational docking studies showed that CDs preferentially bonded to the pocket of laccase on the basal plane rather than the edge through hydrogen bonds and hydrophobic interactions. Electrospray ionization-Fourier transform-ion cyclotron resonance mass spectrometry (ESI-FT-ICR MS) and other characterizations revealed that the phenolic/amino lignins and tannins portions in CDs are susceptible to laccase transformation, resulting in graphitic structure damage and smaller-sized fragments. By using the 13C stable isotope labeling technique, we quantified the uptake and translocation of 13C-CDs by mung bean plants. 13C-CDs (10 mg L-1) accumulated in the root, stem, and leaf were estimated to be 291, 239, and 152 µg g-1 at day 5. We also evidenced that laccase treatment alters the particle size and surface chemistry of CDs, which could facilitate the uptake of CDs by plants and reduce their nanotoxicity to plants.

Mineralization of Few-Layer Graphene Made It Bioavailable in Chlamydomonas reinhardtii.

Numerous studies have emphasized the toxicity of graphene-based nanomaterials to algae, however, the fundamental behavior and processes of graphene in biological hosts, including its transportation, metabolization, and bioavailability, are still not well understood. As photosynthetic organisms, algae are key contributors to carbon fixation and may play an important role in the fate of graphene. This study investigated the biological fate of 14C-labeled few-layer graphene (14C-FLG) in Chlamydomonas reinhardtii (C. reinhardtii). The results showed that 14C-FLG was taken up by C. reinhardtii and then translocated into its chloroplast. Metabolomic analysis revealed that 14C-FLG altered the metabolic profiles (including sugar metabolism, fatty acid, and tricarboxylic acid cycle) of C. reinhardtii, which promoted the photosynthesis of C. reinhardtii and then enhanced their growth. More importantly, the internalized 14C-FLG was metabolized into 14CO2, which was then used to participate in the metabolic processes required for life. Approximately 61.63%, 25.31%, and 13.06% of the total radioactivity (from 14CO2) was detected in carbohydrates, lipids, and proteins of algae, respectively. Overall, these results reveal the role of algae in the fate of graphene and highlight the potential of available graphene in bringing biological effects to algae, which helps to better assess the environmental risks of graphene.

Uptake Route Altered the Bioavailability of Graphene in Misgurnus anguillicaudatus: Comparing Waterborne and Sediment Exposures.

Numerous studies on the bioavailability of graphene-based nanomaterials relate to the water-only exposure route. However, the sediment exposure route should be the most important pathway for benthic organisms to ingest graphene, while to date little work on the bioavailability of graphene in benthic organisms has been explored. In this study, with the help of carbon-14-labeled few-layer graphene (14C-FLG), we quantificationally compared the bioaccumulation, biodistribution, and elimination kinetics of 14C-FLG in loaches via waterborne and sediment exposures. After 72 h of exposure, the accumulated 14C-FLG in loaches exposed via waterborne was 14.28 µg/g (dry mass), which was 3.18 times higher than that (4.49 µg/g) exposed via sediment. The biodistribution results showed that, compared to waterborne exposure, sediment exposure remarkably facilitated the transport of 14C-FLG from the gut into the liver, which made it difficult to be excreted. Although 14C-FLG did not cause significant hepatotoxicity, the disruption of intestinal microbiota homeostasis, immune response, and several key metabolic pathways in the gut were observed, which may be due to the majority of 14C-FLG being accumulated in the gut. Overall, this study reveals the different bioavailabilities of graphene in loaches via waterborne and sediment exposures, which is helpful in predicting its bioaccumulation capability and trophic transfer ability.

Root Hair Apex is the Key Site for Symplastic Delivery of Graphene into Plants.

Uptake kinetics and delivery mechanisms of nanoparticles (NPs) in crop plants need to be urgently understood for the application of nanotechnology in agriculture as delivery systems for eco-friendly nanoagrochemicals. Here, we investigated the uptake kinetics, translocation pathway, and key internalization process of graphene in wheat (Triticum aestivum L.) by applying three specific hydroponic cultivation methods (submerging, hanging, and split-root). Quantification results on the uptake of carbon-14 radiolabeled graphene in each tissue indicated that graphene could enter the root of wheat and further translocate to the shoot with a low delivery rate (<2%). Transmission electron microscopy (TEM) images showed that internalized graphene was transported to adjacent cells through the plasmodesmata, clearly indicating the symplastic pathway of graphene translocation. The key site for the introduction of graphene into root cells for translocation through the symplastic pathway is evidenced to be the apex of growing root hair, where the newly constructed primary cell wall is much thinner. The confirmation of uptake kinetics and delivery mechanisms is useful for the development of nanotechnology in sustainable agriculture, especially for graphene serving as the delivery vector for pesticides, genes, and sensors.

Growth performance and ecological services evaluation of razor clams based on dynamic energy budget model.

Bivalve shellfish aquaculture has been proposed to abate eutrophication and increase carbon sink in integrated multi-trophic aquaculture ecosystems. An individual growth model for razor clams Sinonovacula constricta in an integrated aquaculture pond of Portunus trituberculatus- Penaeus japonicus- S. constricta was constructed based on dynamic energy budget (DEB) theory after parameter measurement and model validation. Goodness-of-fit indices (R-squared, mean difference, and absolute and relative root mean square error) showed that the DEB model accurately reproduced razor clam growth. The growth performance evaluation of razor clams under different environmental conditions showed that warming climate and food shortages inhibited the razor clam growth. The quantification results of ecological service showed that individual razor clam have the potential for nutrient (nitrogen and phosphorus) removal and CO2 fixation, but exhibit a source of CO2 in individual month. The possible applications and contribution of this aquaculture model in China are also discussed, and the assessment results can provide important support for the low carbon bivalve integrated aquaculture.

Inactivation of Laccase by the Attack of As (III) Reaction in Water.

Laccase is a multicopper oxidase containing four coppers as reaction sites, including one type 1, one type 2, and two type 3. We here provide the first experimental data showing that As (III) can be effectively removed from water and transformed to As (V) through reactions mediated by laccase with the presence of oxygen. To this end, the As (III) removal, As (V) yields, total protein, active laccase, and copper concentrations in the aqueous phase were determined, respectively. Additionally, electron paramagnetic resonance spectra and UV-vis spectra were applied to probe possible structural changes of the laccase during the reaction. The data offer the first evidence that laccase can be inactivated by As (III) attack thus leading to the release of type 2 copper. The released copper has no reactivity with the As (III). These findings provide new ideas into a significant pathway likely to master the environmental transformation of arsenite, and advance the understanding of laccase inactivation mechanisms, thus providing a foundation for optimization of enzyme-based processes and potential development for removal and remediation of arsenite contamination in the environment.

Bioaccumulation of 14C-Labeled Graphene in an Aquatic Food Chain through Direct Uptake or Trophic Transfer.

The growing applications of graphene materials warrant a careful evaluation of their environmental fate in aquatic food webs. Escherichia coli (Bacteria), Tetrahymena thermophila (protozoa), Daphnia magna (zooplankton), and Danio rerio (vertebrate) were used to build aquatic food chains to investigate the waterborne uptake and trophic transfer of 14C-labeled graphene. Body burden factor (BBF) and trophic transfer factor (TTF) were analyzed for each organism and food chain to assess the bioaccumulation and biomagnification of graphene. The test organisms have high potential of accumulating graphene via direct uptake from culture medium with log-transformed BBF (log BBF) values of 3.66, 5.1, 3.9, and 1.62 for each organism, respectively. In the food chain from E. coli to T. thermophila, the calculated TTFs of 0.2 to 8.6 indicate the high trophic transfer potential in this aquatic food chain. However, the TTFs calculated for the food chain from T. thermophila to D. magna and from D. magna to D. rerio are much lower than 1, indicating that biomagnification was unlikely to occur in these food chains. Body burden measured for dietary uptake by T. thermophila, D. magna, and D. rerio are higher than that via waterborne exposure in a similar nominal concentration, respectively, indicating that trophic transfer is a nonnegligible route for the bioaccumulation of graphene in organisms.

Bioremediation potential of the hard clam Mercenaria mercenaria as an intensive shrimp aquaculture pond polyculture condidate.

Polyculture practices are important for achieving sustainable aquaculture development. Recently, hard clams polyculture in intensive shrimp ponds has been encouraged because bivalves can consume excess nutrients in aquaculture systems and sequester carbon. To evaluate the bioremediation potential of hard clams polyculture in intensive shrimp ponds, this study built an assessment model based on individual growth models and estimated the potential for nitrogen and phosphorus removal as well as CO2 fixation by hard clams. Firstly, key parameters required for model construction were obtained through field surveys and physiological experiments. Subsequently, an individual growth model for the hard clam Mercenaria mercenaria was developed based on the Dynamic Energy Budget (DEB) theory. Fitting of the growth data indicated that the model accurately replicated the growth patterns of hard clams, with relative root mean square errors of 9.87 % for shell length and 5.02 % for dry tissue weight. Finally, the assessment model for the bioremediation potential of hard clams demonstrated that, over 110 days in the intensive shrimp mariculture pond, the net removal of nitrogen and phosphorus by hard clams were 3.68 kg ha-1 and 0.81 kg ha-1, respectively, and CO2 fixation was 507.00 kg ha-1. These findings suggested that the DEB model is an effective tool for evaluating bivalve ecological remediation potential and can aid in selecting species for sustainable polyculture.

Biological uptake and depuration of radio-labeled graphene by Daphnia magna.

Graphene layers are potential candidates in a large number of applications. However, little is known about their ecotoxicological risks largely as a result of a lack of quantification techniques in complex environmental matrices. In this study, graphene was synthesized by means of graphitization and exfoliation of sandwich-like FePO4/dodecylamine hybrid nanosheets, and (14)C was incorporated in the synthesis. (14)C-labeled graphene was spiked to artificial freshwater and the uptake and depuration of graphene by Daphnia magna were assessed. After exposure for 24 h to a 250 µg/L solution of graphene, the graphene concentration in the organism was nearly 1% of the organism dry mass. These organisms excreted the graphene to clean artificial freshwater and achieved roughly constant body burdens after 24 h depuration periods regardless of the initial graphene exposure concentration. Addition of algae and humic acid to water during the depuration period resulted in release of a significant fraction (>90%) of the accumulated graphene, but some still remained in the organism. Accumulated graphene in adult Daphnia was likely transferred to the neonates. The uptake and elimination results provided here support the environmental risk assessment of graphene and the graphene quantification method is a powerful tool for additional studies.

High temperatures enhance the strength of multiple predator effects in a typical crab-clam system.

Although marine heatwaves pose urgent threats to marine life, our understanding of how these events influence interactions between key species in marine ecosystems is still inadequate. Herein, we examined the behavioral mechanisms by which heat regulates multiple predator effects in different foraging systems that include Asian paddle crabs (Charybdis japonica) and swimming crabs (Portunus trituberculatus) by quantifying their predation and competition at two temperatures. Our results show that non-independent multiple predator effects occurred in the conspecific treatment of Asian paddle crabs and in the interspecific treatment, whereas independent multiple predator effects occurred in the conspecific treatment of swimming crabs. Asymmetrical behavior responses of these crabs to competition and heat triggered divergences in multiple predator effects. High temperatures increased the strength of multiple predator effects but did not alter their types. The reason is that heat negatively impacts predation by enhancing aggressive interactions, outweighing its direct positive effects on predation.

A Dynamic Energy Budget Model for Kuruma Shrimp Penaeus japonicus: Parameterization and Application in Integrated Marine Pond Aquaculture.

Individual growth models can form the basis of population dynamics assessment and ecosystem model construction. In order to provide a basic module for an ecosystem model of an integrated marine aquaculture pond, an individual growth model was constructed for kuruma shrimp (Penaeus japonicus) based on dynamic energy budget (DEB) theory. The model was first parameterized based on a covariation method using the Add-my-Pet (AmP) procedure. The parametric estimation model underestimated the ultimate abdominal length for female shrimp, and the predicted values of other zero-variate parameters were generally consistent with observed values. The relative errors of the predicted and observed values of the univariate data set within three geographical regions showed acceptable goodness of fit. Parameter estimation achieved an overall goodness of fit with a mean relative error of 0.048 and a symmetric mean squared error of 0.066. A DEB model was constructed using the estimated parameters, and the goodness-of-fit indicators (R square, mean bias and absolute and relative root mean square error) showed that the model was able to reproduce the growth of kuruma shrimp in terms of total length and wet weight with high accuracy. The results provide data to support the subsequent development of integrated aquaculture management at the ecosystem level.

Kupffer Cells Degrade 14C-Labeled Few-Layer Graphene to 14CO2 in Liver through Erythrophagocytosis.

The distribution and clearance of graphene materials as drug delivery systems at organ and suborgan levels over the long term remain unclear. Here we compared the fate of 14C-labeled few-layer graphene with different lateral sizes in mice after one intravenous injection for up to 1 year and demonstrated that few-layer graphene mainly accumulated in the liver, and larger graphene can be degraded into 14CO2 by Kupffer cells. The mechanism involves the uptake of graphene by liver cells, larger graphene-induced membrane perturbation of red blood cells, and enhanced erythrophagocytosis by the Kupffer cells, resulting in the degradation of hemoglobin into hemes and a rise in iron concentrations in cells. The increased iron triggered a Fenton reaction to generate the hydroxyl radical, facilitating the degradation of larger graphene into 14CO2. Our findings propose a mechanism for the transformation of graphene that significantly contributes to our understanding of the hepatic fate of graphene in vivo.

Transformation and removal of imidacloprid mediated by silver ferrite nanoparticle facilitated peroxymonosulfate activation in water: Reaction rates, products, and pathways.

Imidacloprid (IMI) is one of the most extensively used chlorinated organic pesticides and its widespread occurrence makes it attract increased public concern and scientific interest. Peroxymonosulfate (PMS) activation has been widely studied for the elimination of organic pollutants from water. But few studies are focused on their heterogeneous catalytic performance towards imidacloprid especially with the presence of silver ferrite nanoparticles (nAgFeO2)-based catalysts. Herein, the catalyst, nAgFeO2, was prepared via a co-precipitation method, and further applied to activate PMS for the removal of imidacloprid (IMI). Our results demonstrated that the prepared nAgFeO2 significantly promoted the activation of PMS for removing IMI, and the removal of IMI followed a pseudo first-order kinetics model with the corresponding nAgFeO2 dosage. Electron paramagnetic resonance (EPR) and quenching tests revealed the singlet oxygen (1O2)-mediated nonradical pathway, instead of hydroxyl radical (•OH) or sulfate radical (SO4•-), played the dominant role in the degradation of IMI. Eight products were identified and the degradation pathways of IMI were proposed. It is postulated that the primary site at the C-1 position of IMI was more easily attacked by the •OH yielding (6-chloropyridin-3-yl) methanol). While the site at the amidine nitrogen (2) of IMI was more likely attacked by the 1O2, and then reacted with •OH to produce 5-hydroxy imidacloprid. Overall, this study provides insights into the mechanisms of nonradical oxidation processes based on PMS for the elimination of pesticides from water, broadening the application of silver ferrite nanoparticles in wastewater treatment.

Uptake of iron oxide nanoparticles inhibits the photosynthesis of the wheat after foliar exposure.

Iron oxide nanoparticles (nFe2O3)-filled materials have been widely employed in various products and their effects on plants have attracted considerable attention because of their potential release into the environment. Currently, numerous studies reporting the influences of iron-bearing nanoparticles on plants are focused on root or seed exposure. However, plants exposed to atmospheric iron-bearing nanoparticles through the leaves and their impacts on plants are still not well understood. This study focused on the uptake, translocation, and effects of foliar exposure of nFe2O3 on wheat seedlings. Wheat seedlings were foliar applied to various concentrations of nFe2O3 (0, 60 and 180 µg per plant) for 1, 7, 14 or 21 d. Our results demonstrated that after exposure for 21 d, the concentrations of Fe in leaves, stems, and roots were 1100, 280 and 160 µg kg-1, respectively. Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), as well as the backscattered electron (BSE) images, revealed the stomatal opening was likely the pathway for nFe2O3 uptake. Analysis of the transfer rate, translocation of Fe from leaves to stems and roots, suggested the involvement of plant Fe regulation processes. Particularly, the antioxidant enzymatic activities and malondialdehyde levels in leaves were modified, which was ascribed to the excessive hydroxyl radical (OH) generated via the Fenton-like reaction mediated by nFe2O3. Finally, the OH facilitated the degradation of chlorophyll, posting a negative impact on the photosynthesis, and thus inhibited the biomass production. These findings are meaningful to understand the fate and physiological effects of atmospheric nFe2O3 in crops.

Adsorption behavior and mechanism of Fe-Mn binary oxide nanoparticles: Adsorption of methylene blue.

Wastewater containing organic dyes has caused worldwide concern. It is thus imperative to develop materials to remove organic dyes from wastewater. In this study, a nano-structured Fe-Mn binary oxide (nFMBO) was synthesized via a facile coprecipitation approach and used for methylene blue (MB) removal from aqueous solution. Characteristic results indicated that the as-prepared nFMBO had a typical wrinkled structure. The adsorption performance of the nFMBO was then investigated by batch experiments. The adsorption kinetics was well fitted to a pseudo-second-order kinetics model, and the adsorption isotherms agreed well with the Langmuir model with a maximum adsorption capacity of 72.32 mg/g at 25 °C. Solution pH was a key factor for adsorption and the absorbent exhibited better removal efficiency for MB in solution with high pH. In addition, it was found that the investigated coexisting anions (CO32-, SO42-, PO33-) did not have a significant influence on MB removal. More importantly, the nFMBO could be easily separated from the water and regenerated by acid elution, and the adsorption efficiency of the nFMBO only decreased to 85.1% of the initial capacity after five adsorption-regeneration cycles. These results indicate that the nFMBO can become an alternative adsorbent for the removal of MB from wastewater.

Non-covalent assembled laccase-graphene composite: Property, stability and performance in beta-blocker removal.

Immobilization of enzymes on carriers have been pursued to make the enzyme stable, reusable and obtaining even better enzyme activity. Due to the highly stable two-dimensional layer structure, large surface area and pore volume, graphene materials were seemed as ideal carrier for enzyme immobilization. In this paper, pristine few layer graphene (FLG) was applied to interact with laccase to synthesize laccase-graphene composite and the results of AFM, FT-IR and adsorption isotherm suggested that laccase was loaded on the FLG with a very high loading dosage (221.1 mg g-1). Based on the measured interaction force and binding type between laccase and graphene, we proposed that the great enzyme loading on FLG is likely due to the non-covalent π-π stacking in addition to the large surface area of FLG. The composite has better stability to the variance of pH and storage temperature than free laccase. The synthesized composite can effectively transform beta-blocker labetalol with an enhanced efficiency, though the possible reaction pathways kept not changing. We further performed molecular simulation study on the crystal structure variation of laccase binding on FLG and proposed that catalytic activity enhancement may be attributed to the more exposure extent of the catalytic center of laccase. In addition, the laccase-graphene composite can be reused more than ten times in catalyzing the labetalol removal.

Biological Uptake, Distribution, and Depuration of Radio-Labeled Graphene in Adult Zebrafish: Effects of Graphene Size and Natural Organic Matter.

The exciting commercial application potential of graphene materials may inevitably lead to their increasing release into the environment where they may pose ecological risks. This study focused on using carbon-14-labeled few-layer graphene (FLG) to determine whether the size of graphene plays a role in its uptake, depuration, and biodistribution in adult zebrafish. After 48 h exposure to larger FLG (L-FLG) at 250 µg/L, the amount of graphene in the organism was close to 48 mg/kg fish dry mass, which was more than 170-fold greater than the body burden of those exposed to the same concentration of smaller FLG (S-FLG). The amount of uptake for both L-FLG and S-FLG increased by a factor of 2.5 and 16, respectively, when natural organic matter (NOM) was added in the exposure suspension. While the L-FLG mainly accumulated in the gut of adult zebrafish, the S-FLG was found in both the gut and liver after exposure with or without NOM. Strikingly, the S-FLG was able to pass through the intestinal wall and enter the intestinal epithelial cells and blood. The presence of NOM increased the quantity of S-FLG in these cells. Exposure to L-FLG or S-FLG also had a significantly different impact on the intestinal microbial community structure.

Graphene Facilitated Removal of Labetalol in Laccase-ABTS System: Reaction Efficiency, Pathways and Mechanism.

The widespread occurrence of the beta-blocker labetalol causes environmental health concern. Enzymatic reactions are highly efficient and specific offering biochemical transformation of trace contaminants with short reaction time and little to none energy consumption. Our experiments indicate that labetalol can be effectively transformed by laccase-catalyzed reaction using 2, 2-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as a mediator, while no significant removal of labetalol can be achieved in the absence of ABTS. A total of three products were identified. It is interesting that the presence of graphene greatly increased the reaction rate while not changed the products. In the presence of 100 µg/L graphene, the pseudo-first-order reaction rate constant was increased ~50 times. We found that the enhancement of graphene is probably attributed to the formation and releasing of ABTS(2+) which has a much greater reactivity towards labetalol when graphene is present. This study provides fundamental information for laccase-ABTS mediated labetalol reactions and the effect of graphene, which could eventually lead to development of novel methods to control beta-blocker contamination.

Comparison of lignin peroxidase and horseradish peroxidase for catalyzing the removal of nonylphenol from water.

Concentrations of aqueous-phase nonylphenol (NP), a well-known endocrine-disrupting chemical, are shown to be reduced effectively via reaction with lignin peroxidase (LiP) or horseradish peroxidase (HRP) and hydrogen peroxide. We systematically assessed their reaction efficiencies at varying conditions, and the results have confirmed that the catalytic performance of LiP toward NP was more efficient than that of HRP under experimental conditions. Mass spectrum analysis demonstrated that polymerization through radical-radical coupling mechanism was the pathway leading to NP transformation. Our molecular modeling with the assistance of ab initio suggested the coupling of NP likely proceeded via covalent bonding between two NP radicals at their unsubstituted carbons in phenolic rings. Data from acute immobilization tests with Daphnia confirm that NP toxicity is effectively eliminated by LiP/HRP-catalyzed NP removal. The findings in this study provide useful information for understanding LiP/HRP-mediated NP reactions, and comparison of enzymatic performance can present their advantages for up-scale applications in water/wastewater treatment.