Liquid metals (LMs) have been used for design of advanced materials by leveraging their unique liquid and electronic properties. Here, we design a series of liquid metal-buffered amorphous molybdenum sulfide nanozymes with different gallium indium ratios, in which Ga75.5In24.5 MoSX nanozymes exhibit the enzyme-like activity, superior to the crystalline MoS2. This has been verified and assigned to the roles of liquid metal: (i) serving as the building template, (ii) phase engineering with catalytic active sites, (iii) electron-rich microenvironment for improving catalytic activity. Besides, the amorphous Ga75.5In24.5 MoSX nanozyme possesses oxidase (OXD)-like and nicotinamide adenine dinucleotide oxidase (NOX)-like activity to achieve multiple-enzyme-like cascade catalytic reactions, disrupting intratumoral redox and metabolism homeostasis. As the example of leveraging liquid metal as a well-designed platform to screen high-performance nanozyme with excellent therapeutic effects, this work highlights the roles of tailored electronic structure and phase engineering for regulation of catalytic activity of nanozymes and may also broaden the application of liquid metal in catalysis and biomedical field.

Multifunctional FeSN nanozyme-doped hydrogel: A promising strategy for antibacterial therapy and wound repair.

Oxidative and reductive reactions are vital processes in cellular metabolism. Like yin and yang in Tai Chi, they are independent and complementary. The non-natural cofactors NCD+ and NCDH exhibit substantial potential for the bio-orthogonal regulation of redox pathways. To provide sufficient driving force, it is crucial to maintain the NCD+ and NCDH ratio and homeostasis. In the Research Article 10.1002/cbic.202500254, Xueying Wang, Zongbao K. Zhao, and co-workers explain how they reshaped the NADH-binding pocket of NADH oxidase to accommodate NCDH, thereby facilitating the traceless regeneration of the oxidized cofactor NCD+ and the selective synthesis of chiral compounds.

Nicotinamide adenine dinucleotide (NAD+) and its reduced form NADH are universal redox cofactors, and thus manipulating NAD(H) supply often leads to unpredictable outcome because of metabolic crosstalk. To overcome intrinsic limitations associated to natural cofactors, this study previously introduces nicotinamide cytosine dinucleotide (NCD+) as a non-natural cofactor for redox biochemistry. While several enzymes have been devised to generate NCDH as driving force at the expense of cheap chemicals for reductive metabolic reactions, it remains inaccessible to generate NCD+ for oxidative reactions. In this study, it engineers an H2O-forming NADH oxidase (EfNOX) from Enterococcus faecalis to favor NCDH. Compared to the wild-type enzyme, the best mutant NADH oxidase (NOX)-KRGT oxidizes NCDH with 14- and 107-fold higher catalytic efficiency and selectivity, respectively. Docking analysis shows that those mutations acquired a narrower cofactor binding cavity and positively charged environment contributing to the preference toward NCDH. Coupling NOX mutants with NCD-favoring phosphite dehydrogenase mutant enables Escherichia coli BW14329 to utilize phosphite as sole phosphorus source for growth. This work provides a traceless and effective tool to convert NCDH into NCD+, which should greatly expand our capacity in developing NCD-linked redox subsystems and further facilitate the implementation of non-natural cofactors in chemical biology and synthetic biology.

High efficacy of camphene and metronidazole combination therapy against Giardia lamblia infection in mice

Single‐atom rhodium mimicking the oxidase and peroxidase for NADH cascade oxidation

Renal interstitial fibrosis (RIF) represents the final pathway in most progressive renal diseases. Curculigoside (CCG), derived from Curculigo Pilosa, affects oxidative stress and inflammation. However, the effects of CCG on RIF remain unclear. This study explored the nephroprotective role of CCG in regulating oxidative stress through the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. Bioinformatic analysis was employed to identify the targets of CCG, elucidate the underlying pathways, and analyze molecular docking results. C57BL/6 mouse models of unilateral ureteral obstruction (UUO) were established to validate the results. Morphologic changes were assessed by pathologic examination, and the expression of proteins associated with renal ferroptosis and fibrosis was analyzed by western blotting. Additionally, the levels of glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and iron were measured. In total, 3,532 differentially expressed genes (DEGs) were identified, comprising 2,290 upregulated and 1,242 downregulated genes. We retrieved 484 ferroptosis-related genes from the ferroptosis regulators (FerrDb) database, identifying 143 DEGs after intersecting with those from the Gene Expression Omnibus Series 217654 dataset (GSE217654). The key identified genes included nicotinamide adenine dinucleotide oxidase 4 (NOX4), activating transcription factor 3 (ATF3), mitogen-activated protein kinase 14 (MAPK14), tissue inhibitor metalloproteinase 1 (TIMP1), and early growth response 1 (EGR1). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that these genes were enriched in oxidative signaling pathways. The results exhibited the docking activity of CCG with related targets. CCG significantly alleviated histopathologic damage, reduced MDA and iron levels, and increased GSH and SOD levels. Protein analysis indicated that CCG alleviated fibrosis and enhanced the protein expression of antioxidants in UUO kidney tissues. CCG activated the Nrf2/HO-1 pathway and reduced UUO-induced ferroptosis. CCG may improve renal fibrosis and mitigate ferroptosis by activating the Nrf2/HO-1 signaling pathway.

Oxygen-boosted fluorinated prodrug hybrid nanoassemblies for bidirectional amplification of breast cancer ferroptosis

Nicotinamide adenine dinucleotide (NADH) oxidase (NOX) is key in converting NADH to NAD+, crucial for various biochemical pathways. However, natural NOXs are costly and unstable. NOX nanozymes offer a promising alternative with potential applications in bio-sensing, antibacterial treatments, anti-aging, and anticancer therapies. This review provides a comprehensive overview of the types, functional mechanisms, biomedical applications, and future research perspectives of NOX nanozymes. It also addresses the primary challenges and future directions in the research and development of NOX nanozymes, underscoring the critical need for continued investigation in this promising area. These challenges include optimizing the catalytic efficiency, ensuring biocompatibility, and achieving targeted delivery and controlled activity within biological systems. Additionally, the exploration of novel materials and hybrid structures holds great potential for enhancing the functional capabilities of NOX nanozymes. Future research directions can involve integrating advanced computational modeling with experimental techniques to better understand the underlying mechanisms and to design more effective nanozyme candidates. Collaborative efforts across disciplines such as nanotechnology, biochemistry, and medicine will be essential to unlock the full potential of NOX nanozymes in future biomedical applications.

Single active Au1O5 clusters for metabolism-inspired sepsis management through immune regulation

Cerium oxide (CeO2-x) performs well in photothermal and catalytic properties due to its abundance of oxygen vacancies. Based on this, we designed a thermosensitive therapeutic nanoplatform to achieve continuous circular drug release in tumor. It can solve the limitation caused by insufficient substrate in the process of tumor treatment. Briefly, CeO2-x and camptothecin (CPT) were wrapped in an agarose hydrogel, which could be melted by the photothermal effect of CeO2-x. At the same time, the local temperature increase provided photothermal treatment, which could induce the apoptosis of tumor cell. After that, CPT was released to damage the DNA in tumor cells to realize chemical treatment. In addition, CPT could active nicotinamide adenine dinucleotide oxidase to react with O2 to increase the intracellular H2O2. After that, the exposed CeO2-x could catalyze H2O2 to generate cytotoxic reactive oxygen species for chemodynamic therapy. More importantly, CeO2-x could catalyze H2O2 to produce O2, which could combine with the catalytic action of CPT to construct a substrate self-cycling nanoenzyme system. Overall, this self-cycling nanoplatform released hypoxia in the tumor microenvironment and built a multimode tumor treatment, which achieved an ideal antitumor affect.

Increasing cellular immunogenicity and reshaping the immune tumor microenvironment (TME) are crucial for antitumor immunotherapy. Herein, this work develops a novel single-atom nanozyme pyroptosis initiator: UK5099 and pyruvate oxidase (POx)-co-loaded Cu-NS single-atom nanozyme (Cu-NS@UK@POx), that not only trigger pyroptosis through cascade biocatalysis to boost the immunogenicity of tumor cells, but also remodel the immunosuppressive TME by targeting pyruvate metabolism. By replacing N with weakly electronegative S, the original spatial symmetry of the Cu-N4 electron distribution is changed and the enzyme-catalyzed process is effectively regulated. Compared to spatially symmetric Cu-N4 single-atom nanozymes (Cu-N4 SA), the S-doped spatially asymmetric single-atom nanozymes (Cu-NS SA) exhibit stronger oxidase activities, including peroxidase (POD), nicotinamide adenine dinucleotide (NADH) oxidase (NOx), L-cysteine oxidase (LCO), and glutathione oxidase (GSHOx), which can cause enough reactive oxygen species (ROS) storms to trigger pyroptosis. Moreover, the synergistic effect of Cu-NS SA, UK5099, and POx can target pyruvate metabolism, which not only improves the immune TME but also increases the degree of pyroptosis. This study provides a two-pronged treatment strategy that can significantly activate antitumor immunotherapy effects via ROS storms, NADH/glutathione/L-cysteine consumption, pyruvate oxidation, and lactic acid (LA)/ATP depletion, triggering pyroptosis and regulating metabolism. This work provides a broad vision for expanding antitumor immunotherapy.

Multi-enzymatic strategies have shown improvement in bioconversion during cofactor regeneration. In this study, purified l-arabinitol 4-dehydrogenase (LAD) and nicotinamide adenine dinucleotide oxidase (Nox) were immobilized via individual, mixed, and sequential co-immobilization approaches on magnetic nanoparticles, and were evaluated to enhance the conversion of l-arabinitol to l-xylulose. Initially, the immobilization of LAD or Nox on the nanoparticles resulted in a maximum immobilization yield and relative activity of 91.4% and 98.8%, respectively. The immobilized enzymes showed better pH and temperature profiles than the corresponding free enzymes. Furthermore, co-immobilization of these enzymes via mixed and sequential methods resulted in high loadings of 114 and 122 mg/g of support, respectively. Sequential co-immobilization of these enzymes proved more beneficial for higher conversion than mixed co-immobilization because of better retaining Nox residual activity. Sequentially co-immobilized enzymes showed a high relative conversion yield with broader pH, temperature, and storage stability profiles than the controls, along with high reusability. To the best of our knowledge, this is the first report on the mixed or sequential co-immobilization of LAD and Nox on magnetic nanoparticles for l-xylulose production. This finding suggests that selecting a sequential co-immobilization strategy is more beneficial than using individual or mixed co-immobilized enzymes on magnetic nanoparticles for enhancing conversion applications.

Covalent coupling of functionalized outer membrane vesicles (OMVs) to gold nanoparticles

The thermostability of thermophilic enzymes makes them ideal for use in biotechnology and medical science research. The overexpression of heterogeneous genes from thermophiles, such as the mesophilic Deinococcus geothermalis, in Escherichia coli is a genetic engineering technique for producing stable and useful proteins. In our previous study, 1- or 2-base expansion of the Shine-Dalgarno sequence, which is a ribosome-binding site in mRNA, was effective for the overexpression of the nicotinamide adenine dinucleotide oxidase gene from Deinococcus. geothermalis in Escherichia coli. In the present study, we examined the effect of expanding the Shine-Dalgarno sequence of the malate dehydrogenase gene and three genes from the aldehyde dehydrogenase family. Our results revealed that a 1- or 2-base expansion of the Shine-Dalgarno sequence was effective for the overexpression of all genes from Deinococcus geothermalis in Escherichia coli. However, the effects of the expansion of Shine-Dalgarno sequence from 3 to 5 bases were different between these genes.

Yogurt is made by fermenting milk with 2 lactic acid bacteria, Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus. To comprehensively understand the protocooperation mechanism between S. thermophilus and L. bulgaricus in yogurt fermentation, we examined 24 combinations of cocultures comprising 7 fast- or slow-acidifying S. thermophilus strains with 6 fast- or slow-acidifying L. bulgaricus strains. Furthermore, 3 NADH oxidase (Nox)-deficient mutants (Δnox) and one pyruvate formate-lyase deficient mutant (ΔpflB) of S. thermophilus were used to evaluate the factor that determines the acidification rate of S. thermophilus. The results revealed that the acidification rate of S. thermophilus monoculture determined the yogurt fermentation rates, despite the coexistence of L. bulgaricus, whose acidification rate was either fast or slow. Significant correlation was found between the acidification rate of S. thermophilus monoculture and the amount of formate production. Result using ΔpflB showed that the formate was indispensable for the acidification of S. thermophilus. Moreover, results of the Δnox experiments revealed that formate production required Nox activity, which not only regulated dissolved oxygen, but also the redox potential. The Nox provided the large decrease in redox potential required by pyruvate formate-lyase to produce formate. A highly significant correlation was found between formate accumulation and Nox activity in S. thermophilus. In conclusion, the formate production ability provided by the action of Nox activity determines the acidification rate of S. thermophilus, and consequently, regulates yogurt coculture fermentation.

Deep Brain Stimulation for the Management of AIFM1-Related Disabling Tremor: A Case Series

Anaerobic conversion rate of phenol to methane was low due to its biological toxicity. In this study, the coupling of granular activated carbon (GAC) and exogenous hydrogen (EH) could enhance greatly methane production of phenol anaerobic digestion, and the metagenomic was firstly used to analyze its potential mechanism. The results indicated that a mass of syntrophic acetate-oxidizing bacteria and hydrogen-utilizing methanogens were enriched on the GAC surface, and SAO-HM pathway has become the dominant pathway. The energy transfer analysis implied that the abundance of adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide (NADH) oxidase increased. Furthermore, direct interspecies electron transfer (DIET) was formed by promoting type IV e-pili between Methanobacterium and Syntrophus, thereby improving the interspecies electron transfer efficiency. The dominant SAO-HM pathway was induced and DIET was formed, which was the internal mechanism of the coupling of GAC and EH to enhance anaerobic biotransformation of phenol.

Immobilized multienzyme systems aregaining momentum in appliedbiocatalysis; however, the coimmobilization of several enzymes onone carrier is still challenging. In this work, we exploited a heterofunctionalsupport activated with three different chemical functionalities toimmobilize a wide variety of different enzymes. This support is basedon agarose microbeads activated with aldehyde, amino, and cobalt chelatemoieties that allow a fast and irreversible immobilization of enzymes,enhancing the thermostability of most of the heterogeneous biocatalysts(up to 21-fold higher than the soluble one). Furthermore, this trifunctionalsupport serves to efficiently coimmobilize a multienzyme system composedof an alcohol dehydrogenase, a reduced nicotinamide adenine dinucleotide(NADH) oxidase, and a catalase. The confined multienzymatic systemdemonstrates higher performance than its free counterpart, achievinga total turnover number (TTN) of 1 × 105 during fivebatch consecutive cycles. We envision this solid material as a platformfor coimmobilizing multienzyme systems with enhanced properties tocatalyze stepwise biotransformations.

Nanozyme catalytic therapy triggered by tumor-specific endogenous stimuli is an emerging tumor therapy that attracts wide attention. However, the current therapeutic efficacy of nanozyme catalytic therapy is severely limited by the catalytic efficiency of nanozymes and the concentration of endogenous reaction substrates. Herein, a novel and efficient IrN5 single-atom (IrN5 SA) nanozyme is developed with multiple enzyme-like catalytic activities. Due to the synergistic effect of central Ir single-atom and axial N coordination, IrN5 SA exhibits better enzymatic catalytic performance than IrN4 SA. At tumor sites, IrN5 SA can generate a large amount of reactive oxygen species (ROS) through oxidase (OXD)-like and peroxidase (POD)-like catalytic activities. Moreover, IrN5 SA can also generate O2 and hydrogen peroxide (H2 O2 ) through catalase (CAT)-like and nicotinamide adenine dinucleotide (NADH) oxidase (NOX)-like catalytic activities, realizing the efficient nanozyme catalytic therapy in a substrate-cycle manner. Additionally, IrN5 SA can effectively break the intracellular NADH/NAD+ cycle balance by mimicking NOX, and then cooperate with fatty acid synthase cerulenin (Cer) to interfere with the energy metabolism homeostasis of tumor cells. Consequently, the designed IrN5 SA/Cer nanoagent can disrupt redox and metabolic homeostasis in the tumor region through an enzyme-mimicking cascade reaction, effectively overcoming the shortcomings of current nanozyme catalytic therapy.