Data-Driven Evolutionary Design of Multienzyme-like Nanozymes.

Multienzyme-like nanozymes are nanomaterials with multiple enzyme-like activities and are the focus of nanozyme research owing to their ability to facilitate cascaded reactions, leverage synergistic effects, and exhibit environmentally responsive selectivity. However, multienzyme-like nanozymes exhibit varying enzyme-like activities under different conditions, making them difficult to precisely regulate according to the design requirements. Moreover, individual enzyme-like activity in a multienzyme-like activity may accelerate, compete, or antagonize each other, rendering the overall activity a complex interplay of these factors rather than a simple sum of single enzyme-like activity. A theoretically guided strategy is highly desired to accelerate the design of multienzyme-like nanozymes. Herein, nanozyme information was collected from 4159 publications to build a nanozyme database covering element type, element ratio, chemical valence, shape, pH, etc. Based on the clustering correlation coefficients of the nanozyme information, the material features in distinct nanozyme classifications were reorganized to generate compositional factors for multienzyme-like nanozymes. Moreover, advanced methods were developed, including the quantum mechanics/molecular mechanics method for analyzing the surface adsorption and binding energies of substrates, transition states, and products in the reaction pathways, along with machine learning algorithms to identify the optimal reaction pathway, to aid the evolutionary design of multienzyme-like nanozymes. This approach culminated in creating CuMnCo7O12, a highly active multienzyme-like nanozyme. This process is named the genetic-like evolutionary design of nanozymes because it resembles biological genetic evolution in nature and offers a feasible protocol and theoretical foundation for constructing multienzyme-like nanozymes.

Peptidoglycan biosynthesis-associated enzymatic kinetic characteristics and ß-lactam antibiotic inhibitory effects of different Streptococcus pneumoniae penicillin-binding proteins.

Penicillin-binding proteins (PBPs) include transpeptidases, carboxypeptidases, and endopeptidases for biosynthesis of peptidoglycans in the cell wall to maintain bacterial morphology and survival in the environment. Streptococcus pneumoniae expresses six PBPs, but their enzymatic kinetic characteristics and inhibitory effects on different ß-lactam antibiotics remain poorly understood. In this study, all the six recombinant PBPs of S. pneumoniae displayed transpeptidase activity with different substrate affinities (Km = 1.56-9.11 mM) in a concentration-dependent manner, and rPBP3 showed a greater catalytic efficiency (Kcat = 2.38 s-1) than the other rPBPs (Kcat = 3.20-7.49 × 10-2 s-1). However, only rPBP3 was identified as a carboxypeptidase (Km = 8.57 mM and Kcat = 2.57 s-1). None of the rPBPs exhibited endopeptidase activity. Penicillin and cefotaxime inhibited the transpeptidase and carboxypeptidase activity of all the rPBPs but imipenem did not inhibited the enzymatic activities of rPBP3. Except for the lack of binding of imipenem to rPBP3, penicillin, cefotaxime, and imipenem bound to all the other rPBPs (KD = 3.71-9.35 × 10-4 M). Sublethal concentrations of penicillin, cefotaxime, and imipenem induced a decrease of pneumococcal pbps-mRNA levels (p < 0.05). These results indicated that all six PBPs of S. pneumoniae are transpeptidases, while only PBP3 is a carboxypeptidase. Imipenem has no inhibitory effect on pneumococcal PBP3. The pneumococcal genes for encoding endopeptidases remain to be determined.

Effect of heterogenous dopant and high temperature pulse excitation on ozone sensing behavior of In2O3 nanostructures and an image recognition method coupled to ozone sensing array.

Ground-level ozone (O3) is a primary air pollutant with potential adverse impacts on human health and ecosystems. Aiming to detect O3 concentration and develop efficient O3 sensing materials, sensing behavior of heterogenous cation (Fe3+, Sn4+ and Sb5+) doped In2O3 nanostructures was investigated. The incorporation of these cations modulated the electronic structure of semiconductor oxides, affecting the density of chemisorbed oxygen species and reactive sites. From O3 sensing results, Fe3+ doped In2O3 based sensors featuring saturated resistance curves in O3 gas, demonstrated fast sensing speed and qualified detection threshold (20 ppb). In contrast, Sn4+ and Sb5+ doped counterparts exhibited non-saturated sensing curves, resulting in slower response/recovery speed. As a proof-of-concept, these optimized sensors were integrated as the sensor array. Coupled to the image recognition technique, this sensor array could successfully discriminate O3 and NOx. That is, through the tailored combination of material modulation and sensor array, this study paves a novel approach for highly sensitive and selective O3 detection.

Multienzyme-Like Nanozyme Encapsulated Ocular Microneedles for Keratitis Treatment.

Keratitis, an inflammation of the cornea caused by bacterial or fungal infections, is one of the leading causes of severe visual disability and blindness. Keratitis treatment requires both the prevention of infection and the reduction of inflammation. However, owing to their limited therapeutic functions, in addition to the ocular barrier, existing conventional medications are characterized by poor efficacy and low bioavailability, requiring high dosages or frequent topical treatment, which represents a burden on patients and increases the risk of side effects. In this study, manganese oxide nanocluster-decorated graphdiyne nanosheets (MnOx/GDY) are developed as multienzyme-like nanozymes for the treatment of infectious keratitis and loaded into hyaluronic acid and polymethyl methacrylate-based ocular microneedles (MGMN). MGMN not only exhibits antimicrobial and anti-inflammatory effects owing to its multienzyme-like activities, including oxidase, peroxidase, catalase, and superoxide dismutase mimics but also crosses the ocular barrier and shows increased bioavailability via the microneedle system. Moreover, MGMN is demonstrated to eliminate pathogens, prevent biofilm formation, reduce inflammation, alleviate ocular hypoxia, and promote the repair of corneal epithelial damage in in vitro, ex vivo, and in vivo experiments, thus providing a better therapeutic effect than commercial ophthalmic voriconazole, with no obvious microbial resistance or cytotoxicity.

Ultrasound-Augmented Multienzyme-like Nanozyme Hydrogel Spray for Promoting Diabetic Wound Healing.

Treatment of diabetic foot ulcers (DFU) needs to reduce inflammation, relieve hypoxia, lower blood glucose, promote angiogenesis, and eliminate pathogenic bacteria, but the therapeutic efficacy is greatly limited by the diversity and synergy of drug functions as well as the DFU microenvironment itself. Herein, an ultrasound-augmented multienzyme-like nanozyme hydrogel spray was developed using hyaluronic acid encapsulated l-arginine and ultrasmall gold nanoparticles and Cu1.6O nanoparticles coloaded phosphorus doped graphitic carbon nitride nanosheets (ACPCAH). This nanozyme hydrogel spray possesses five types of enzyme-like activities, including superoxide dismutase (SOD)-, catalase (CAT)-, glucose oxidase (GOx)-, peroxidase (POD)-, and nitric oxide synthase (NOS)-like activities. The kinetics and reaction mechanism of the sonodynamic/sonothermal synergistic enhancement of the SOD-CAT-GOx-POD/NOS cascade reaction of ACPCAH are fully investigated. Both in vitro and in vivo tests demonstrate that this nanozyme hydrogel spray can be activated by the DFU microenvironment to reduce inflammation, relieve hypoxia, lower blood glucose, promote angiogenesis, and eliminate pathogenic bacteria, thus accelerating diabetic wound healing effectively. This study highlights a competitive approach based on multienzyme-like nanozymes for the development of all-in-one DFU therapies.

Optical polarization properties of TPP2MnBr4 perovskite crystal adjusted by the self-trapping state.

Low-dimensional networked organic-inorganic hybrid metal halide crystal has become an emerging hotspot material due to its opportunities and advantages in the development of white-light-emitting diodes. Therefore, its photoluminescence (PL) mechanism is important. Herein, we study the PL behavior of columniform TPP2MnBr4 crystals using multi-spectroscopy. The temperature-dependent PL data show that the PL of the TPP2MnBr4 crystal originates from the recombination of a self-trapping exciton. A polarization-dependent PL test suggests that the self-trapping exciton is anisotropic, which indicates that the distribution of self-trapping states is sensitive to the orientation of the crystal axis. Space-resolved PL spectroscopy shows that the anisotropy of PL gradually weakens along the orientation of the columniform crystal, which has a longer relaxation distance than traditional light-wave-guiding behavior. Thus, anisotropy of PL can exist before it disappears in the crystal. Our results elucidate the PL mechanism of low-dimensional networked organic-inorganic hybrid metal halide crystals and provide a foundation for advanced optical polarization devices based on them.

Metal-organic frameworks derived inverse/normal bimetallic spinel oxides toward the selective VOCs and H2S sensing.

As the typical toxic and hazardous gases, volatile organic compounds (VOCs) and hydrogen sulfide (H2S) pose a threat to the environment and human health. The demand for real-time detection of VOCs and H2S gases is growing in many application to protect human health and air quality. Therefore, it is essential to develop advance sensing materials for the construction of effective and reliable gas sensors. Herein, bimetallic spinel ferrites with different metal ions (MFe2O4, M = Co, Ni, Cu and Zn) were designed by using metal-organic frameworks as templates. The evaluation of cation substitution on crystal structures (inverse/normal spinel structure) and electrical properties (n/p type and band gap) is systematically discussed. The results indicate that p-type NiFe2O4 and n-type CuFe2O4 nanocubes with inverse spinel structure exhibit high response and great selectivity towards acetone (C3H6O) and H2S, respectively. Moreover, the two sensors also display the detection limits as low as 1 ppm (C3H6O) and 0.5 ppm (H2S), which are far below the threshold values of 750 ppm to acetone and 10 ppm to H2S for 8 h exposure set by American Conference of Governmental Industrial Hygienists (ACGIH). The finding provides new possibilities for the design of high-performance chemical sensors, which display tremendous potential for practical applications.

Boosting charge separation in graphdiyne quantum dots/hollow tubular carbon nitride heterojunction for water pollutant degradation.

Non-desirable solar energy absorption and poor charge transfer efficiency are two problems that limit the peroxymonosulfate (PMS) photocatalytic techniques. Herein, a metal-free boron-doped graphdiyne quantum dot (BGDs) modified hollow tubular g-C3N4 photocatalyst (BGD/TCN) was synthesized to activate PMS and achieved effective space separation of carriers for degradation of bisphenol A. With 0.5 mM PMS, the degradation rate of bisphenol A (20 ppm) was 0.0634 min-1, 3.7-fold higher than that of TCN itself. The roles of BGDs in the distribution of electrons and photocatalytic property were well identified by experiments and density functional theory (DFT) calculations. The possible degradation intermediate products of bisphenol A were monitored by mass spectrometer and demonstrated to be nontoxic using ecological structure activity relationship modeling (ECOSAR). Finally, this newly-designed material was successfully applied in actual water bodies, which further renders its promising prospect for actual water remediation.

Competition of Carrier Kinetics Contributes to Amplified Spontaneous Emission in Quasi-2D/3D (PBA)2MAn-1PbnBr3n+1 Thin Films under Strip Light Mode.

Quasi-2D halide perovskites have potential in lasing due to their amplified spontaneous emission (ASE) properties. The ASE of (PBA)2MAn-1PbnBr3n+1 thin films has been confirmed by photoluminescence (PL) testing using stripe light excitation (SLE). The ASE threshold decreases with decreasing environmental temperature (TE) or increasing number of inorganic layers (n). Using the transient absorption technique, the Auger recombination and the cooling process of the high-activity carrier are accelerated with the decrease of n or TE. A new ASE mechanism is proposed where high-activity carriers directly emit photons under photon perturbation from adjacent sites, leading to the accumulation and amplification of emitted photons only in the SLE region for ASE to occur. In addition, the reduction of n promotes light scattering between nano-thin layers, which supports a rapid increase in the ASE signal after the ASE threshold is crossed.

Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY.

SPINDLY (SPY) in Arabidopsis thaliana is a novel nucleocytoplasmic protein O-fucosyltransferase (POFUT), which regulates diverse developmental processes. Sequence analysis indicates that SPY is distinct from ER-localized POFUTs and contains N-terminal tetratricopeptide repeats (TPRs) and a C-terminal catalytic domain resembling the O-linked-N-acetylglucosamine (GlcNAc) transferases (OGTs). However, the structural feature that determines the distinct enzymatic selectivity of SPY remains unknown. Here we report the cryo-electron microscopy (cryo-EM) structure of SPY and its complex with GDP-fucose, revealing distinct active-site features enabling GDP-fucose instead of UDP-GlcNAc binding. SPY forms an antiparallel dimer instead of the X-shaped dimer in human OGT, and its catalytic domain interconverts among multiple conformations. Analysis of mass spectrometry, co-IP, fucosylation activity, and cryo-EM data further demonstrates that the N-terminal disordered peptide in SPY contains trans auto-fucosylation sites and inhibits the POFUT activity, whereas TPRs 1-5 dynamically regulate SPY activity by interfering with protein substrate binding.

Ultrasound activatable microneedles for bilaterally augmented sono-chemodynamic and sonothermal antibacterial therapy.

Chemodynamic therapy (CDT) employs Fenton catalysts to kill bacteria by converting hydrogen peroxide (H2O2) into toxic hydroxyl radical (•OH). Among them, Fenton-type metal peroxide nanoparticles fascinate nanomaterials with intriguing physiochemical properties, but research on this antibacterial agent is still in its infancy. Herein, a distinct CuO2/TiO2 heterostructure constituted of ultrasmall copper peroxide (CuO2) nanoclusters and sonosensitized ultrathin oxygen vacancy-rich porous titanium oxide (OV-TiO2) nanosheets was developed and was incorporated into microneedles for bilaterally augmented sono-chemodynamic and sonothermal antibacterial therapy. Engineering CuO2 nanoclusters on the surface of TiO2 nanosheets not only endows the Fenton catalytic activity for sono-chemodynamic therapy (SCDT), but also improves the sonodynamic and sonothermal performance of TiO2 by narrowing the bandgap of TiO2 and suppressing the recombination of electron-hole pairs. The high efficacy of this CuO2/TiO2 integrated microneedle (CTMN) patch was systematically demonstrated both in vitro and in vivo with the eliminating rate >99.9999% against multidrug resistant (MDR) pathogens in 5 min as well as accelerated wound tissue healing. This work highlights a promisingly new and efficient strategy for the development of sonosensitive and chemoreactive nanomedicine for non-antibiotic therapies. STATEMENT OF SIGNIFICANCE: Feton-type metal peroxides, a novel nanomaterial with self-supplied oxygen and hydrogen peroxide, can achieve effective antimicrobial activity in vitro. However, there is a lack of effective nanomaterial delivery systems and suitable means for in vivo activation/enhancement of antimicrobial activity during bacterial infected skin wound treatment. In this study, we designed and prepared efficient ultrasound activable microneedles that effectively addressed the deficiencies mentioned above and established a new paradigm for efficient utilization of metal peroxide nanomaterials and ultrasound based strategies. Noticeably, copper peroxide nanoclusters/oxygen vacancy-rich porous titanium oxide nanosheets (CuO2/TiO2) integrated microneedle (CTMN) patch combines advantages of both sono-chemodynamic and sonothermal antibacterial therapy, achieving one of the most instant and effective antibacterial efficacy (>99.9999% in 5 min) in vivo reported till now.

Machine Learning Guided Discovery of Superoxide Dismutase Nanozymes for Androgenetic Alopecia.

Androgenetic alopecia (AGA) is a common form of hair loss, which is mainly caused by oxidative stress induced dysregulation of hair follicles (HF). Herein, a highly efficient manganese thiophosphite (MnPS3) based superoxide dismutase (SOD) mimic was discovered using machine learning (ML) tools. Remarkably, the IC50 of MnPS3 is 3.61 µg·mL-1, up to 12-fold lower than most reported SOD-like nanozymes. Moreover, a MnPS3 microneedle patch (MnMNP) was constructed to treat AGA that could diffuse into the deep skin where HFs exist and remove excess reactive oxygen species. Compared with the widely used minoxidil, MnMNP exhibits higher ability on hair regeneration, even at a reduced frequency of application. This study not only provides a general guideline for the accelerated discovery of SOD-like nanozymes by ML techniques, but also shows a great potential as a next generation approach for rational design of nanozymes.

Activity enhancement of a photo-generated carrier in CsPbBr3 nanocrystals improved by Cd element.

Doping Cd element into perovskite materials is an effective strategy to improve the photoelectric property. However, the further discussion for carrier dynamic behavior in perovskites affected by Cd element remains not sufficient. In this research letter, based on steady and transient spectroscopy, it is found that adding Cd element into CsPbBr3 nanocrystals can enhance the activity of photo-generated carriers and accompany with the optimization of crystal structure. The former improves the carrier heating effect, which makes carrier keep high temperature for a long time and accelerate the bimolecular and the Auger recombination simultaneously. The latter can restrict the monomolecular recombination through passivating the defect states. Finally, they together improve the photoluminescence characteristics of the Cd doped CsPbBr3 nanocrystals and make them exhibit a huge potential in the fields of optoelectronics or photo-catalysis.

A Dual Sensing Platform for Human Exhaled Breath Enabled by Fe-MIL-101-NH2 Metal-Organic Frameworks and its Derived Co/Ni/Fe Trimetallic Oxides.

Limited by the insufficient active sites and the interference from breath humidity, designing reliable gas sensing materials with high activity and moisture resistance remains a challenge to analyze human exhaled breath for the translational application of medical diagnostics. Herein, the dual sensing and cooperative diagnosis is achieved by utilizing metal-organic frameworks (MOFs) and its derivative. The Fe-MIL-101-NH2 serves as the quartz crystal microbalance humidity sensing layer, which exhibits high selectivity and rapid response time (16 s/15 s) to water vapor. Then, the Co2+ and Ni2+ cations are further co-doped into Fe-MIL-101-NH2 host to obtain the derived Co/Ni/Fe trimetallic  oxides (CoNiFe-MOS-n). The chemiresistive CoNiFe-MOS-n sensor displays the high sensitivity (560) and good selectivity to acetone, together with a lower original resistance compared with Fe2 O3 and NiFe2 O4 . Moreover, as a proof-of-concept application, synergistic integration of Fe-MIL-101-NH2 and derived CoNiFe-MOS-n is carried out. The Fe-MIL-101-NH2 is applied as moisture sorbent materials, which realize a sensitivity compensation of CoNiFe-MOS-n sensors for the detection of acetone (biomarker gas of diabetes). The findings provide an insight for effective utilization of MOFs and the derived materials to achieve a trace gas detection in exhaled breath analysis.

Ultrathin Graphdiyne/Graphene Heterostructure as a Robust Electrochemical Sensing Platform.

Graphdiyne (GDY) has been considered as an appealing electrode material for electrochemical sensing because of its alkyne-rich structure and high degrees of π-conjugation, which shows great affinity to heavy metal ions and pollutant molecules via d-π and π-π interactions. However, the low surface area and poor conductivity of bulk GDY limit its electrochemical performance. Herein, a two-dimensional ultrathin GDY/graphene (GDY/G) nanostructure was synthesized and used as an electrode material for electrochemical sensing. Graphene plays the role of an epitaxy template for few-layered GDY growth and conductive layers. The formed few-layered GDY with a high surface area possesses abundant affinity sites toward heavy metal ions (Cd2+, Pb2+) and toxic molecules, for example, nitrobenzene and 4-nitrophenol, via d-π and π-π interactions, respectively. Moreover, hemin as a key part of the enzyme catalytic motif was immobilized on GDY/G via π-π interactions. The artificial enzyme mimic hemin/GDY/G-modified electrode exhibited promising ascorbic acid and uric acid detection performance with excellent sensitivity and selectivity, a good linear range, and reproducibility. More importantly, real sample detection and the feasibility of this electrochemical sensor as a wearable biosensor were demonstrated.

Biosensors for rapid detection of bacterial pathogens in water, food and environment.

Conventional techniques (e.g., culture-based method) for bacterial detection typically require a central laboratory and well-trained technicians, which may take several hours or days. However, recent developments within various disciplines of science and engineering have led to a major paradigm shift in how microorganisms can be detected. The analytical sensors which are widely used for medical applications in the literature are being extended for rapid and on-site monitoring of the bacterial pathogens in food, water and the environment. Especially, within the low-resource settings such as low and middle-income countries, due to the advantages of low cost, rapidness and potential for field-testing, their use is indispensable for sustainable development of the regions. Within this context, this paper discusses analytical methods and biosensors which can be used to ensure food safety, water quality and environmental monitoring. In brief, most of our discussion is focused on various rapid sensors including biosensors and microfluidic chips. The analytical performances such as the sensitivity, specificity and usability of these sensors, as well as a brief comparison with the conventional techniques for bacteria detection, form the core part of the discussion. Furthermore, we provide a holistic viewpoint on how future research should focus on exploring the synergy of different sensing technologies by developing an integrated multiplexed, sensitive and accurate sensors that will enable rapid detection for food safety, water and environmental monitoring.

Nanoscale Bimetallic AuPt-Functionalized Metal Oxide Chemiresistors: Ppb-Level and Selective Detection for Ozone and Acetone.

As the most widely used gas sensors, metal oxide semiconductor (MOS)-based chemiresistors have been facing great challenges in achieving ppb-level and selective detection of the target gas. The rational design and employment of bimetallic nanocatalysts (NCs) are expected to address this issue. In this work, the well-shaped and monodispersed AuPt NCs (diameter ≈ 9 nm) were functionalized on one-dimensional (1D) In2O3 nanofibers (NFs) to construct efficient gas sensors. The sensor demonstrated dual-selective and ppb-level detection for ozone (O3) and acetone (C3H6O) at different optimal working temperatures. For the possible application exploitation, a circuit was designed to monitor O3 concentration and provide warnings when the concentration safety limit (50 ppb) was exceeded. Moreover, simulated exhaled breath measurements were also carried out to diagnose diabetes through C3H6O concentration. The selective detection for O3 and C3H6O was further analyzed by principal component analysis (PCA). The drastically enhanced sensing performances were attributed to the synergistic catalytic effect of AuPt NCs. Both the "spillover effect" and the Schottky barrier at the interfaces of AuPt NCs and In2O3 NFs promoted the sensing processes of O3 and C3H6O.

Piezoelectric Activatable Nanozyme-Based Skin Patch for Rapid Wound Disinfection.

Nanozymes are promising new-generation antibacterial agents owing to their low cost, high stability, broad-spectrum activity, and minimal antimicrobial resistance. However, the inherent low catalytic activity of nanozymes tends to limit their antibacterial efficacy. Herein, a heterostructure of zinc oxide nanorod@graphdiyne nanosheets (ZnO@GDY NR) with unparallel piezocatalytic enzyme mimic activity is reported, which concurrently possesses intrinsic peroxidase-like activity and strong piezoelectric responses and effectively promotes the decomposition of hydrogen peroxide (H2O2) and generation of reactive oxygen species under ultrasound irradiation. Moreover, this piezocatalytic nanozyme exhibits almost 100% antibacterial efficacy against multidrug-resistant pathogens involving methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa in vitro and in vivo. In addition, a piezoelectric activatable nanozyme-based skin patch is developed for rapid skin wound disinfections with satisfactory hemocompatibility and cytocompatibility. This work not only sheds light on the development of an innovative piezoelectric activatable nanozyme-based skin patch for rapid wound disinfection but also provides new insights on the engineering of piezocatalytic nanozymes for nanozyme antibacterial therapy.

Nonmetal Graphdiyne Nanozyme-Based Ferroptosis-Apoptosis Strategy for Colon Cancer Therapy.

Ferroptosis-apoptosis, a new modality of induced cell death dependent on reactive oxygen species, has drawn tremendous attention in the field of nanomedicine. A metal-free ferroptosis-apoptosis inducer was reported based on boron and nitrogen codoped graphdiyne (BN-GDY) that possesses efficient glutathione (GSH) depletion capability and concurrently induces ferroptosis by deactivation of GSH-dependent peroxidases 4 (GPX4) and apoptosis by downregulation of Bcl2. The high catalytic activity of BN-GDY is explicated by both kinetic experiments and density functional theory (DFT) calculations of Gibbs free energy change during hydrogen peroxide (H2O2) decomposition. In addition, a unique sequence Bi-Bi mechanism is discovered, which is distinct from the commonly reported ping-pong Bi-Bi mechanism of most peroxidase mimics and natural enzymes. We anticipate that this nonmetal ferroptosis-apoptosis therapeutic concept by carbon-based nanomaterials would provide proof-of-concept evidence for nanocatalytic medicines in cancer therapy.