Ammonium release in synthetic and human urine by a urease immobilized nanoconstruct.

In this work, we have studied the ability of urease immobilized on glutaraldehyde crosslinked chitosan coated magnetic iron oxide nanoparticles (Urease/GA/CS/MIONPs), for the hitherto unreported comparative hydrolysis of urea in synthetic (SUr) and real human urine (HUr). The prepared Urease/GA/CS/MIONPs were characterized by a combination of Fourier transform infrared spectroscopy (FTIR), field emission-scanning-electron-microscopy (FESEM), energy dispersive X-ray spectroscopy (EDX) and dynamic light scattering (DLS). The nanoconstructs display the highest ammonium ion liberation post-urea hydrolysis in 1/20 or 1/24-fold dilutions of SUr and HUr, respectively. The optimum activity of immobilized urease is observed at pH 7, and the nanoconstructs facilitate efficient urea-hydrolysis till at least 45 °C. Kinetic analysis of the immobilized urease shows km and vmax of 14.81 mM, 12.36 mM, and 18.55 µM min-1 and 10.10 µM min-1, towards SUr and HUr, respectively. The magnetization of the immobilized urease is suitable for reuse across multiple cycles of urea hydrolysis in SUr and HUr. The robust performance of Urease/GA/CS/MIONPs in SUr and HUr is promising for generating ammonium as a useable source of nitrogen from human urine, and underscores the suitability of SUr as a urine mimic for such interventions.

G-quadruplexes in MTOR and induction of autophagy.

G-quadruplex (G4) structures have emerged as singular therapeutic targets for cancer and neurodegeneration. Autophagy, a crucial homeostatic mechanism of the cell, is often dysregulated in neurodegenerative diseases and cancers. We used QGRS mapper to identify 470 G4 sequences in MTOR, a key negative regulator of autophagy. We sought to identify a functional context by leveraging the effect of G4-targeting ligands on MTOR G4 sequences. The effect of Bis-4,3, a G4 selective dimeric carbocyanine dye, was compared with the known G4-stabilizing activity of the porphyrin, TMPyP4 in HeLa and SHSY-5Y cells. Our results show that treatment with G4-selective ligands downregulates MTOR RNA and mTOR protein expression levels. This is the first report describing G4 motifs in MTOR. This study indicates a possible role of G4 stabilizing ligands in induction of autophagy by downregulation of mTOR levels, albeit not precluding MTOR independent pathways.

Banana Peel Derived Chitosan-Grafted Biocomposite for Recovery of NH4+ and PO43.

Biomass-derived adsorbents afford accessible and inexpensive harvesting of nitrogen and phosphorus from wastewater sources. Human urine is widely accepted as a rich source of nitrogen and phosphorus. However, direct use of urine in agriculture is untenable because of its unpleasant smell, pathogen contamination, and pharmaceutical residues. In this work, we have grafted chitosan onto dried and crushed banana peel (DCBP) to generate the biocomposite DCBP/Ch. A combination of FTIR, TGA, XRD, FESEM, EDX, and NMR analyses were used to characterize DCBP/Ch and reveal condensation-aided covalent conjugation between O-H functionalities of DCBP and chitosan. The adsorption performance of DCBP/Ch toward NH4+ and PO43- is in sync with its attractive surface porosity, elevated crystallinity, and thermostability. The maximum adsorption capacity of DCBP/Ch toward NH4+/PO43- was estimated as 42.16/15.91 mg g-1 at an operating pH of 7/4, respectively, and ranks highly when compared to previously reported bioadsorbents. DCBP/Ch performs admirably when tested on artificial urine. While nitrogen and phosphorus harvesting from human urine using single techniques has been reported previously, this is the first report of a single adsorbent for recovery of NH4+ and PO43-. The environmental compatibility, ease of preparation, and economic viability of DCBP/Ch present it as an attractive candidate for deployment in waste channels.

An MXene-Grafted Terpolymer Hydrogel for Adsorptive Immobilization of Toxic Pb(II) and Post-Adsorption Application of Metal Ion Hydrogel.

Toxic metal ions present in industrial waste, such as Pb(II), introduce deleterious effects on the environment. Though the adsorptive removal of Pb(II) is widely reported, there is a dearth of research on the suitable utilization and disposal of the Pb(II)-adsorbed adsorbent. In this work, an MXene-grafted terpolymer (MXTP) hydrogel has been designed for the adsorption of Pb(II) under ambient conditions of pH and temperature. The hydrogel MXTP was synthesized by facile one-pot polymerization in aqueous solvent, and the detailed structural characterization of terpolymer (TP), MXTP, and Pb(II)-loaded MXTP, i.e., Pb(II)-MXTP, was carried out by a combination of proton nuclear magnetic resonance (1H NMR), Fourier-transform infrared (FTIR), X-ray photoelectron spectroscopy (XPS), X-ray diffractometric (XRD), thermogravimetric/differential thermogravimetric (TG/ DTG), and field emission scanning electron microscopic (FESEM) analyses. The specific capacitance and conductivities of Pb(II)-MXTP were studied with cyclic voltammetry (CV) and electrical impedance spectroscopy (EIS), which unambiguously indicate successful post-adsorption application. The specific capacitance of MXTP decreased after Pb(II) adsorption, whereas the conductivity increased significantly after Pb(II) adsorption, showing that MXTP can be successfully deployed as a solid electrolyte/anode after Pb(II) adsorption. This study covers the synthesis of a novel MXene-grafted terpolymer hydrogel for adsorptive exclusion of Pb(II) and assessment of the as-adsorbed Pb(II)-loaded hydrogel as a solid electrolyte/anode material and is the first demonstration of such post-adsorptive application.

Bacteria-derived topologies of Cu2O nanozymes exert a variable antibacterial effect.

The ability of bacteria to facilitate fabrication of nanomaterials has been adapted towards bacterial sensing applications. In this work, we fabricate spherical, cubic and truncated octahedron topologies of Cu2O nanoparticles via E. coli-facilitated redox reaction in an electrochemical setup. The Cu2O nanoparticles exhibit cytochrome c oxidase-like activity with the spherical topology displaying higher catalytic rate compared to the other geometries. The topology-dependent catalytic behavior of Cu2O nanoparticles has not been reported previously. The Cu2O nanozymes also display E. coli killing activity in a topology-correlated manner. The E. coli mediated redox reaction in an electrochemical setup is being reported for the first time for synthesis of different topologies of Cu2O which also exert a variable antibacterial effect.

In Silico Identification of Potential Quadruplex Forming Sequences in LncRNAs of Cervical Cancer.

Long non-coding RNAs (lncRNAs) have emerged as auxiliary regulators of gene expression influencing tumor microenvironment, metastasis and radio-resistance in cancer. The presence of lncRNA in extracellular fluids makes them promising diagnostic markers. LncRNAs deploy higher-order structures to facilitate a complex range of functions. Among such structures, G-quadruplexes (G4s) can be detected or targeted by small molecular probes to drive theranostic applications. The in vitro identification of G4 formation in lncRNAs can be a tedious and expensive proposition. Bioinformatics-driven strategies can provide comprehensive and economic alternatives in conjunction with suitable experimental validation. We propose a pipeline to identify G4-forming sequences, protein partners and biological functions associated with dysregulated lncRNAs in cervical cancer. We identified 17 lncRNA clusters which possess transcripts that can fold into a G4 structure. We confirmed in vitro G4 formation in the four biologically active isoforms of SNHG20, MEG3, CRNDE and LINP1 by Circular Dichroism spectroscopy and Thioflavin-T-assisted fluorescence spectroscopy and reverse-transcriptase stop assay. Gene expression data demonstrated that these four lncRNAs can be potential prognostic biomarkers of cervical cancer. Two approaches were employed for identifying G4 specific protein partners for these lncRNAs and FMR2 was a potential interacting partner for all four clusters. We report a detailed investigation of G4 formation in lncRNAs that are dysregulated in cervical cancer. LncRNAs MEG3, CRNDE, LINP1 and SNHG20 are shown to influence cervical cancer progression and we report G4 specific protein partners for these lncRNAs. The protein partners and G4s predicted in lncRNAs can be exploited for theranostic objectives.

Use of Single-Molecule Plasmon-Enhanced Fluorescence to Investigate Ligand Binding to G-Quadruplex DNA.

Single-molecule measurements are crucial for studying the interactions between G-quadruplex (GQ) DNA and ligands, as they provide higher resolution and sensitivity compared to those of bulk measurements. In this study, we employed plasmon-enhanced fluorescence to investigate the real-time interaction between the cationic porphyrin ligand TmPyP4 and different topologies of telomeric GQ DNA at the single-molecule level. By analyzing the time traces of the fluorescence bursts, we extracted dwell times for the ligand. For parallel telomeric GQ DNA, the dwell time distribution followed a biexponential fit, yielding mean dwell times of 5.6 and 18.6 ms. For the antiparallel topology of human telomeric GQ DNA, plasmon-enhanced fluorescence of TmPyP4 was observed, with dwell time distributions following a single-exponential fit and a mean dwell time of 5.9 ms. Our approach allows the nuances of GQ-ligand interactions to be captured and holds promise for studying weakly emitting GQ ligands at the single-molecule level.

Chimeric nanoparticles for targeting mitochondria in cancer cells.

Mitochondrial dysfunction is implicated in myriad diseases, including cancer. Subsequently, targeting mitochondrial DNA (mt-DNA) in cancer cells has emerged as an unorthodox strategy for anti-cancer therapy. However, approaches targeting only one component of the mitochondrial "central dogma" can be evaded by cancer cells through various mechanisms. To address this, herein, we have engineered mitochondria-targeting cholesterol-based chimeric nanoparticles (mt-CNPs) consisting of cisplatin, camptothecin, and tigecycline, which can simultaneously impair mt-DNA, mitochondrial topoisomerase I (mt-Top1), and mitochondrial ribosomes. mt-CNPs were characterized as being positively charged, spherical in shape, and 187 nm in diameter. Confocal microscopy confirmed that mt-CNPs efficiently localized into the mitochondria of A549 lung cancer cells within 6 h, followed by mitochondrial morphology damage and the subsequent generation of reactive oxygen species (ROS). mt-CNPs showed remarkable cancer-cell killing abilities compared to free-drug combinations in A549 (lung), HeLa (cervical), and MCF7 (breast) cancer cells. These mitochondria-targeting lipidic chimeric nanoparticles could be explored further to impair multiple targets in mitochondria, helping researchers to gain an understanding of mitochondrial translational and transcriptional machinery and to develop new strategies for cancer therapy.

Intracellular Bacterial Targeting by a Thiazolyl Benzenesulfonamide and Octaarginine Peptide Complex.

A brominated thiazolyl benzenesulfonamide (BTB) derivative is conjugated with the cell-penetrating peptide octaarginine (R8) in an effort to construct innovative antibacterial products. The noncovalent complex of BTB and R8 is characterized by Fourier transform infrared (FTIR) spectroscopy, which indicates hydrogen bonding between the two constituents. Attachment of the peptide moiety renders aqueous solubility to the hydrophobic benzenesulfonamide drug and bestows bactericidal activity. Confocal imaging in conjunction with dye probes shows successful clearance of intracellular Staphylococcus aureus bacteria by the BTB-R8 complex. Scanning electron micrographs and studies with a set of fluorescent dyes suggest active disruption of the bacterial cell membrane by the BTB-R8 complex. In contrast, the complex of BTB with octalysine (K8) fails to cause membrane damage and displays a modest antibacterial effect. A complex of BTB with the water-soluble hydrophilic polymer poly(vinylpyrrolidone) (PVP) does not display any antibacterial effect, indicating the distinctive role of the cell-penetrating peptide (CPP) R8 in the cognate complex. The leakage of the encapsulated dye from giant unilamellar vesicles upon interaction with the BTB-R8 complex further highlights the membrane activity of the complex, which cannot be accomplished by bare sulfonamide alone. This work broadens the scope of use of CPPs with respect to eliciting antibacterial activity and potentially expands the limited arsenal of membrane-targeting antibiotics.

Design and development of 5-(4H)-oxazolones as potential inhibitors of human carbonic anhydrase VA: towards therapeutic management of diabetes and obesity.

Inhibitors of carbonic anhydrase (CAIs) hold promise for addressing various diseases, including cancer, diabetes, and other metabolic syndromes. CAV is the only isoform present in the mitochondria and is considered a potential drug target for obesity. In this work, we have developed C2, and C4 substituted oxazole-5(4H)-one derivatives as a new scaffold for the selective inhibition of human carbonic anhydrase VA (hCAVA). Synthesized compounds were characterized by 1H NMR, 13C NMR, and LC-MS mass spectrometry and subsequently evaluated for in vitro hCAVA inhibition. Two compounds, 4 and 5 showed a considerably higher binding affinity for hCAVA in comparison to the hCAII. Further, cell-based studies showed that these compounds decrease the expression of CAVA and GLUT4 in adipocytes and non-toxic to HEK293 cells. The present work opens a platform for the use of oxazole-5(4H)-ones and holds promise for further refinement of potent and selective hCAVA inhibitors.Communicated by Ramaswamy H. Sarma.

Stimuli Responsive, Programmable DNA Nanodevices for Biomedical Applications.

Of the multiple areas of applications of DNA nanotechnology, stimuli-responsive nanodevices have emerged as an elite branch of research owing to the advantages of molecular programmability of DNA structures and stimuli-responsiveness of motifs and DNA itself. These classes of devices present multiples areas to explore for basic and applied science using dynamic DNA nanotechnology. Herein, we take the stake in the recent progress of this fast-growing sub-area of DNA nanotechnology. We discuss different stimuli, motifs, scaffolds, and mechanisms of stimuli-responsive behaviours of DNA nanodevices with appropriate examples. Similarly, we present a multitude of biological applications that have been explored using DNA nanodevices, such as biosensing, in vivo pH-mapping, drug delivery, and therapy. We conclude by discussing the challenges and opportunities as well as future prospects of this emerging research area within DNA nanotechnology.

Assessing G4-Binding Ligands In Vitro and in Cellulo Using Dimeric Carbocyanine Dye Displacement Assay.

G-quadruplexes (G4) are the most actively studied non-canonical secondary structures formed by contiguous repeats of guanines in DNA or RNA strands. Small molecule mediated targeting of G-quadruplexes has emerged as an attractive tool for visualization and stabilization of these structures inside the cell. Limited number of DNA and RNA G4-selective assays have been reported for primary ligand screening. A combination of fluorescence spectroscopy, AFM, CD, PAGE, and confocal microscopy have been used to assess a dimeric carbocyanine dye B6,5 for screening G4-binding ligands in vitro and in cellulo. The dye B6,5 interacts with physiologically relevant DNA and RNA G4 structures, resulting in fluorescence enhancement of the molecule as an in vitro readout for G4 selectivity. Interaction of the dye with G4 is accompanied by quadruplex stabilization that extends its use in primary screening of G4 specific ligands. The molecule is cell permeable and enables visualization of quadruplex dominated cellular regions of nucleoli using confocal microscopy. The dye is displaced by quarfloxin in live cells. The dye B6,5 shows remarkable duplex to quadruplex selectivity in vitro along with ligand-like stabilization of DNA G4 structures. Cell permeability and response to RNA G4 structures project the dye with interesting theranostic potential. Our results validate that B6,5 can serve the dual purpose of visualization of DNA and RNA G4 structures and screening of G4 specific ligands, and adds to the limited number of probes with such potential.

Nanomaterials for Agricultural and Ecological Defense Applications: Active Agents and Sensors.

The world we live in today is overpopulated with an unprecedented number of people competing for fewer and fewer precious resources. The struggle to efficiently steward and manage these resources is a global problem in need of concrete and urgent solutions. Nanomaterials have driven innovation in diverse industrial sectors including military, aviation, electronic, and medical among others. Nanoscale materials possess unique surfaces and exquisite opto-electronic properties that make them uniquely suited to environmental, biological, and ecological defense applications. A tremendous upsurge of research activity in these areas is evident from the exponential increase in publications worldwide. Here we review recent applications of nanomaterials toward soil health and management, abiotic and biotic stress management, plant defense, delivery of the RNA Interference (RNAi), plant growth, manufacture of agro-products, and ecological investigations related to farming. For example, nanomaterial constructs have been used to counter environmental stresses and in plant defense and disease diagnosis. Nanosensor chemistries have been developed to monitor water quality and measure specific pollutant levels. Specific nanomaterials such as silver, iron oxide, and zinc oxide proffer protection to plants from pathogens. This review describes progress in nanomaterial-based agricultural and ecological defense and seeks to identify factors that would enable their wider commercialization and deployment. This article is categorized under: Diagnostic Tools > Biosensing Toxicology and Regulatory Issues in Nanomedicine > Toxicology of Nanomaterials Diagnostic Tools > Diagnostic Nanodevices.

Nanoparticle-Mediated Routing of Antibiotics into Mitochondria in Cancer Cells.

In recent years, antibiotics have emerged as alternative medicines in cancer therapy due to their capability of mitochondrial dysfunction in cancer cells. However, antibiotics render collateral damage in noncancerous cells by targeting mitochondrial transcription and translational machinery. To address this, herein, we have engineered three different mitochondria-targeted cationic antibiotic (tigecycline)-loaded nanoparticles from cholesterol conjugates. Dynamic light scattering and electron microscopy confirmed the spherical morphology and a less than 200 nm hydrodynamic diameter for these nanoparticles. The triphenylphosphine-coated tigecycline-loaded nanoparticle (Mito-TPP-Tig-NP) was shown to be homed into the mitochondria of A549 lung cancer cells compared to the other cationic nanoparticles. These Mito-TPP-Tig-NPs indeed triggered mitochondrial morphology damage and generation of reactive oxygen species (ROS). All the mitochondria-targeted tigecycline-loaded nanoparticles showed improved cancer cell killing ability in A549 and HeLa cervical cancer cells compared to free tigecycline. Moreover, Mito-TPP-Tig-NPs showed much less toxicity toward noncancerous human embryonic kidney cells (HEK293) compared to free tigecycline. These antibiotic-loaded mitochondria-targeted nanoparticles can open up an avenue toward anticancer therapy.

Emergent antibacterial activity of N-(thiazol-2-yl)benzenesulfonamides in conjunction with cell-penetrating octaarginine.

Hybrid antimicrobials that combine the effect of two or more agents represent a promising antibacterial therapeutic strategy. In this work, we have synthesized N-(4-(4-(methylsulfonyl)phenyl)-5-phenylthiazol-2-yl)benzenesulfonamide derivatives that combine thiazole and sulfonamide, groups with known antibacterial activity. These molecules are investigated for their antibacterial activity, in isolation and in complex with the cell-penetrating peptide octaarginine. Several of the synthesized compounds display potent antibacterial activity against both Gram-negative and Gram-positive bacteria. Compounds with 4-tert-butyl and 4-isopropyl substitutions exhibit attractive antibacterial activity against multiple strains. The isopropyl substituted derivative displays low MIC of 3.9 µg mL-1 against S. aureus and A. xylosoxidans. The comparative antibacterial behaviour of drug-peptide complex, drug alone and peptide alone indicates a distinctive mode of action of the drug-peptide complex, that is not the simple sum total of its constituent components. Specificity of the drug-peptide complex is evident from comparison of antibacterial behaviour with a synthetic intermediate-peptide complex. The octaarginine-drug complex displays faster killing-kinetics towards bacterial cells, creates pores in the bacterial cell membranes and shows negligible haemolytic activity towards human RBCs. Our results demonstrate that mere attachment of a hydrophobic moiety to a cell penetrating peptide does not impart antibacterial activity to the resultant complex. Conversely, the work suggests distinctive modes of antibiotic activity of small molecules when used in conjunction with a cell penetrating peptide.

Small Molecule Soluble Epoxide Hydrolase Inhibitors in Multitarget and Combination Therapies for Inflammation and Cancer.

The enzyme soluble epoxide hydrolase (sEH) plays a central role in metabolism of bioactive lipid signaling molecules. The substrate-specific hydrolase activity of sEH converts epoxyeicosatrienoic acids (EETs) to less bioactive dihydroxyeicosatrienoic acids. EETs exhibit anti-inflammatory, analgesic, antihypertensive, cardio-protective and organ-protective properties. Accordingly, sEH inhibition is a promising therapeutic strategy for addressing a variety of diseases. In this review, we describe small molecule architectures that have been commonly deployed as sEH inhibitors with respect to angiogenesis, inflammation and cancer. We juxtapose commonly used synthetic scaffolds and natural products within the paradigm of a multitarget approach for addressing inflammation and inflammation induced carcinogenesis. Structural insights from the inhibitor complexes and novel strategies for development of sEH-based multitarget inhibitors are also presented. While sEH inhibition is likely to suppress inflammation-induced carcinogenesis, it can also lead to enhanced angiogenesis via increased EET concentrations. In this regard, sEH inhibitors in combination chemotherapy are described. Urea and amide-based architectures feature prominently across multitarget inhibition and combination chemotherapy applications of sEH inhibitors.

DNA-Functionalized Nanoparticles for Targeted Biosensing and Biological Applications.

Nanoscale systems have increasingly been used in biomedical applications, enhancing the demand for the development of biomolecule-functionalized nanoparticles for targeted applications. Such designer nanosystems hold great prospective to refine disease diagnosis and treatment. To completely investigate their potential for bioapplications, nanoparticles must be biocompatible and targetable toward explicit receptors to guarantee particular detecting, imaging, and medication conveyance in complex organic milieus, for example, living cells, tissues, and organisms. We present recent works that explore enhanced biocompatibility and biorecognition of nanoparticles functionalized with DNA and different DNA entities such as aptamers, DNAzymes, and aptazymes. We sum up the methods utilized in the amalgamation of complex nanostructures, survey the significant types of multifunctional nanoparticles that have been developed in the course of recent years, and give a perceptual vision of the significant field of nanomedicine. The field of DNA-functionalized nanoparticles holds an incredible guarantee in rising biomedical zones, for example, multimodal imaging, theranostics, and picture-guided treatments.

Design and Development of Novel Urea, Sulfonyltriurea, and Sulfonamide Derivatives as Potential Inhibitors of Sphingosine Kinase 1.

Sphingosine kinase 1 (SphK1) is one of the well-studied drug targets for cancer and inflammatory diseases. Recently discovered small-molecule inhibitors of SphK1 have been recommended in cancer therapeutics; however, selectivity and potency of first-generation inhibitors are great challenge. In search of effective SphK1 inhibitors, a set of small molecules have been designed and synthesized bearing urea, sulfonylurea, sulfonamide, and sulfonyltriurea groups. The binding affinity of these inhibitors was measured by fluorescence-binding assay and isothermal titration calorimetry. Compounds 1, 5, 6, and 7 showed an admirable binding affinity to the SphK1 in the sub-micromolar range and significantly inhibited SphK1 activity with admirable IC50 values. Molecular docking studies revealed that these compounds fit well into the sphingosine binding pocket of SphK1 and formed significant number of hydrogen bonds and van der Waals interactions. These molecules may be exploited as potent and selective inhibitors of SphK1 that could be implicated in cancer therapeutics after the required in vivo validation.

Nanobiocatalyst facilitated aglycosidic quercetin as a potent inhibitor of tau protein aggregation.

Polyphenols have been suggested as potential therapeutic agents for the treatment of amyloidogenic diseases. In this work, we evaluate quercetin-rich onion extract for its ability to inhibit tau fibrillization. Considering the presence of polyphenols in multiple glycosidic and aglycosidic forms, a nanobiocatalyst-mediated approach has been used to extract quercetin from onion skins. The nanobiocatalysts facilitate greater release of quercetin compared to the use of free enzymes. Atomic force microscopy and fluorescence microscopy show that quercetin possesses a novel inhibitory character on tau-fibril aggregation. In contrast, quercetin-diglucoside does not have an inhibitory effect. Molecular Dynamics simulations reveal conformational changes in tau protein upon interaction with quercetin due to specific hydrogen bonding and hydrophobic interactions. The resulting conformational stability of tau monomer reduces propensity of the protein to aggregate. The ability of quercetin to inhibit tau fibrillization expands the paradigm for application of bioactive polyphenols.

Investigation of nanoparticle immobilized cellulase: nanoparticle identity, linker length and polyphenol hydrolysis.

Cellulase containing nanobiocatalysts have been useful as an extraction tool based on their ability to disrupt plant cell walls. In this work, we investigate the effect of nanoparticle composition and chemical linkage towards immobilized cellulase activity. Cellulase nanoconstructs have been prepared, characterized and compared for their loading efficiencies with standard assays and enzyme kinetics and correlate well with the cognate loading efficiencies. Application of the cellulase-immobilized nanoparticles on onion skins results in release of a distinctive composition of polyphenols. The aglycosidic form of quercetin is the dominant product of onion skin hydrolysis affected by cellulase nanobiocatalysts. Chitosan-coated iron oxide nanoparticles with APTES-conjugated cellulase are found to be most effective for polyphenol release and for transformation of glycosidic to aglycosidic form of quercetin. These results shed light on the activity of immobilized cellulase beyond their role in cell wall disruption and are important for the practical application of cellulase nanobiocatalysts.