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A simple abiological host-guest system demonstrates racemase activity with catalytic rate enhancements of 10 4 without employing traditional functional groups. Cooperative weak interactions enhanced through shape-complementarity between the catalyst active site and the reaction transition state drive this activity, as proposed by Pauling for enzymes. In analogy to the Jencks' concept of catalytic antibodies, it is shown that a hapten resembling the planar transition state of the bowl inversion acts as a potent inhibitor of this catalytic process. In contrast, no substrate/product inhibition is detected, and a relatively weak binding of the substrate is observed ( K a ≈ 10 2 M -1 at 293 K). This simple box-and-bowl system demonstrates that shape selectivity arising from cooperative dispersive forces suffices for the emergence of a catalytic system with an enzyme-like thermodynamic profile.

Disulfide bonds play essential roles in the folding of secretory and plasma membrane proteins in the endoplasmic reticulum (ER). In eukaryotes, protein disulfide isomerase (PDI) is an enzyme catalyzing the disulfide bond formation and isomerization in substrates. The Arabidopsis (Arabidopsis thaliana) genome encodes diverse PDIs including structurally distinct subgroups PDI-L and PDI-M/S. It remains unclear how these AtPDIs function to catalyze the correct disulfide formation. We found that one Arabidopsis ER oxidoreductin-1 (Ero1), AtERO1, can interact with multiple PDIs. PDI-L members AtPDI2/5/6 mainly serve as an isomerase, while PDI-M/S members AtPDI9/10/11 are more efficient in accepting oxidizing equivalents from AtERO1 and catalyzing disulfide bond formation. Accordingly, the pdi9/10/11 triple mutant exhibited much stronger inhibition than pdi1/2/5/6 quadruple mutant under dithiothreitol treatment, which caused disruption of disulfide bonds in plant proteins. Furthermore, AtPDI2/5 work synergistically with PDI-M/S members in relaying disulfide bonds from AtERO1 to substrates. Our findings reveal the distinct but overlapping roles played by two structurally different AtPDI subgroups in oxidative protein folding in the ER., (© American Society of Plant Biologists 2021. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Objective: Human chorionic membrane extracts (CMEs) from placenta are known to be a natural biomaterial for bone regeneration, with their excellent osteogenic efficacy on osteoblasts. However, little is known about the regulatory mechanism involved., Methods and Results: We have shown the in vitro and in vivo bone-forming ability of CME using human osteoblasts and bone defect animal models, suggesting that CME greatly enhances osteogenesis by providing an osteoconductive environment for the osteogenesis of osteoblasts. Proteomic analysis revealed that CME contained several osteogenesis-related stimulators such as osteopontin, osteomodulin, Thy-1, netrin 4, retinol-binding protein and DJ-1. Additionally, 23 growth factors/growth factor-related proteins were found in CME, which may trigger mitogen-activated protein kinase (MAPK) signalling as a specific cellular signalling pathway for osteogenic differentiation. Microarray analysis showed four interaction networks (chemokine, Wnt signalling, angiogenesis and ossification), indicating the possibility that CME can promote osteogenic differentiation through a non-canonical Wnt-mediated CXCL signalling-dependent pathway., Conclusions: The results of this study showed the function and mechanism of action of CME during the osteogenesis of osteoblasts and highlighted a novel strategy for the use of CME as a biocompatible therapeutic material for bone regeneration., (© 2021 The Authors. Cell Proliferation published by John Wiley & Sons Ltd.)

Although a monoclonal antibody targeting the multifunctional ectoenzyme CD38 is an FDA-approved drug, few small molecule inhibitors exist for this enzyme that catalyzes inter alia the formation and metabolism of the N 1-ribosylated, Ca 2+ -mobilizing, second messenger cyclic adenosine 5'-diphosphoribose (cADPR). N 1-Inosine 5'-monophosphate ( N 1-IMP) is a fragment directly related to cADPR. 8-Substituted- N 1-IMP derivatives, prepared by degradation of cyclic parent compounds, inhibit CD38-mediated cADPR hydrolysis more efficiently than related cyclic analogues, making them attractive for inhibitor development. We report a total synthesis of the N 1-IMP scaffold from adenine and a small initial compound series that facilitated early delineation of structure-activity parameters, with analogues evaluated for inhibition of CD38-mediated hydrolysis of cADPR. The 5'-phosphate group proved essential for useful activity, but substitution of this group by a sulfonamide bioisostere was not fruitful. 8-NH 2 - N 1-IMP is the most potent inhibitor (IC 50 = 7.6 μM) and importantly HPLC studies showed this ligand to be cleaved at high CD38 concentrations, confirming its access to the CD38 catalytic machinery and demonstrating the potential of our fragment approach.

This work introduces a novel way to obtain catalytically competent oxyferryl species for two different dye-decolorizing peroxidases (DyPs) in the absence of H 2 O 2 or any other peroxide by simply applying a reductive electrochemical potential under aerobic conditions. UV-vis and resonance Raman spectroscopies show that this method yields long-lived compounds II and I for the DyPs from Bacillus subtilis (BsDyP; Class I) and Pseudomonas putida (PpDyP; Class P), respectively. Both electrochemically generated high valent intermediates are able to oxidize ABTS at both acidic and alkaline pH. Interestingly, the electrocatalytic efficiencies obtained at pH 7.6 are very similar to the values recorded for regular catalytic ABTS/H 2 O 2 assays at the optimal pH of the enzymes, ca. 3.7. These findings pave the way for the design of DyP-based electrocatalytic reactors operable in an extended pH range without the need of harmful reagents such as H 2 O 2 .

In this report, chitosan/zinc oxide (CS/ZnO) nanocomposite was synthesized using Sida acuta and assessed their antibacterial and photocatalytic properties. The formation of CS/ZnO nanocomposite was preliminary confirmed by colour change and UV-visible spectroscopy. The crystalline peaks related to CS and ZnO in CS/ZnO nanocomposite were demonstrated by XRD. Morphological analysis through FE-SEM and TEM showed a rod like appearance for ZnO NPs and agglomerated grains with rod shaped morphology was observed for the CS/ZnO nanocomposite. The peaks around 400-800 cm -1 in the IR spectrum of nanocomposite indicated the vibrations of metal-oxygen (ZnO), whereas bands at 1659 cm -1 and 1546 cm -1 indicated the presence of amine groups, which confirms the CS in the synthesized CS/ZnO nanocomposite. The CS/ZnO nanocomposite exhibited remarkable growth inhibition activity against B. subtilis and E. coli with 22 ± 0.3 and 16.5 ± 0.5 mm zone of inhibitions. In addition, CS/ZnO nanocomposite treated cotton fabrics also exhibited antibacterial activity against B. subtilis and E. coli. Furthermore, the ZnO NPs and nanocomposite showed time depended photodegradation activity and revealed 76% and 91% decomposition of CR under sunlight irradiation. In conclusion, our study revealed that the functionalization of biopolymer CS to the inorganic ZnO enhances the bio and catalytic properties., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Neurolysin (Nln) is a recently recognized endogenous mechanism functioning to preserve the brain from ischemic injury. To further understand the pathophysiological function of this peptidase in stroke and other neurologic disorders, the present study was designed to identify small molecule activators of Nln. Using a computational approach, the structure of Nln was explored, which was followed by docking and in silico screening of ∼140,000 molecules from the National Cancer Institute Developmental Therapeutics Program database. Top ranking compounds were evaluated in an Nln enzymatic assay, and two hit histidine-dipeptides were further studied in detail. The identified dipeptides enhanced the rate of synthetic substrate hydrolysis by recombinant (human and rat) and mouse brain-purified Nln in a concentration-dependent manner (micromolar A 50 and A max ≥ 300%) but had negligible effect on activity of closely related peptidases. Both dipeptides also enhanced hydrolysis of Nln endogenous substrates neurotensin, angiotensin I, and bradykinin and increased efficiency of the synthetic substrate hydrolysis (V max /K m ratio) in a concentration-dependent manner. The dipeptides and competitive inhibitor dynorphin A (1-13) did not affect each other's affinity for Nln, suggesting differing nature of their respective binding sites. Lastly, drug affinity responsive target stability (DARTS) and differential scanning fluorimetry (DSF) assays confirmed concentration-dependent interaction of Nln with the activator molecule. This is the first study demonstrating that Nln activity can be enhanced by small molecules, although the peptidic nature and low potency of the activators limit their application. The identified dipeptides provide a chemical scaffold to develop high-potency, drug-like molecules as research tools and potential drug leads. SIGNIFICANCE STATEMENT: This study describes discovery of two molecules that selectively enhance activity of peptidase Nln-a newly recognized cerebroprotective mechanism in the poststroke brain. The identified molecules will serve as a chemical scaffold for development of drug-like molecules to further study Nln and may become lead structures for a new class of drugs. In addition, our conceptual and methodological framework and research findings might be used for other peptidases and enzymes, the activation of which bears therapeutic potential., (Copyright © 2021 by The American Society for Pharmacology and Experimental Therapeutics.)

Polysaccharides are biopolymers composed of simple sugars like glucose, galactose, mannose, fructose, etc. The major natural sources for the production of polysaccharides include plants and microorganisms. In the present work, four bacterial and two fungal polysaccharides (PS or EPS) were used for the modification and preservation of Pycnoporus sanguineus cellobiose dehydrogenase (CDH) activity. It was found that the presence of polysaccharide preparations clearly enhanced the stability of cellobiose dehydrogenase compared to the control value (4 °C). The highest stabilization effect was observed for CDH modified with Rh110EPS. Changes in the optimum pH in the samples of CDH incubated with the chosen polysaccharide modifiers were evidenced as well. The most significant effect was observed for Rh24EPS and Cu139PS (pH 3.5). Cyclic voltammetry used for the analysis of electrochemical parameters of modified CDH showed the highest peak values after 30 days of incubation with polysaccharides at 4 °C. In summary, natural polysaccharides seem to be an effective biotechnological tool for the modification of CDH activity to increase the possibilities of its practical applications in many fields of industry., (© 2021. The Author(s).)

Ligninolytic enzymes, including laccase, manganese peroxidase, and dye-decolorizing peroxidase (DyP), have attracted much attention in the degradation of mycotoxins. Among these enzymes, the possible degradation pathway of mycotoxins catalyzed by DyP is not yet clear. Herein, a DyP-encoding gene, St DyP, from Streptomyces thermocarboxydus 41291 was identified, cloned, and expressed in Escherichia coli BL21/pG-Tf2. The recombinant St DyP was capable of catalyzing the oxidation of the peroxidase substrate 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), phenolic lignin compounds 2,6-dimethylphenol, and guaiacol, non-phenolic lignin compound veratryl alcohol, Mn 2+ , as well as anthraquinone dye reactive blue 19. Moreover, St DyP was able to slightly degrade zearalenone (ZEN). Most importantly, we found that St DyP combined the catalytic properties of manganese peroxidase and laccase, and could significantly accelerate the enzymatic degradation of ZEN in the presence of their corresponding substrates Mn 2+ and 1-hydroxybenzotriazole. Furthermore, the biological toxicities of the main degradation products 15-OH-ZEN and 13-OH-ZEN-quinone might be remarkably removed. These findings suggested that DyP might be a promising candidate for the efficient degradation of mycotoxins in food and feed.

In this article, a zirconia-based nano-catalyst (Nano-ZrO 2 ), with intermolecular C-N bond formation for the synthesis of various benzimidazole-fused heterocycles in a concise method is reported. The robustness of this reaction is demonstrated by the synthesis of a series of benzimidazole drugs in a one-pot method. All synthesized materials were characterized using 1 HNMR, 13 CNMR, and LC-MS spectroscopy as well as microanalysis data. Furthermore, the synthesis of nano-ZrO 2 was processed using a standard hydrothermal technique in pure form. The crystal structure of nano-ZrO 2 and phase purity were studied, and the crystallite size was calculated from XRD analysis using the Debye-Scherrer equation. Furthermore, the antimicrobial activity of the synthesized benzimidazole drugs was evaluated in terms of Gram-positive, Gram-negative, and antifungal activity, and the results were satisfactory.

Rapid degradation/destruction of chemical warfare agents, especially ones containing a phosphorous-fluorine bond, is of notable interest due to their extreme toxicity and typically rapid rate of human incapacitation. Recent studies of the hydrolytic destruction of a key nerve agent simulant, dimethyl 4-nitrophenylphosphate (DMNP), catalyzed by Zr 6 -based metal-organic frameworks (MOFs), have suggested deactivation of the active sites due to inhibition by the products as the reaction progresses. In this study, the interactions of two MOFs, NU-1000 and MOF-808, and two hydrolysis products, dimethyl phosphate (DMP) and ethyl methyl phosphonate (EMP), from the hydrolysis of the simulant (DMNP) and nerve agent ethyl methylphosphonofluoridate (EMPF), resembling the hydrolysis degradation product of the G-series nerve agent, Sarin (GB), have been investigated to deconvolute the effect of product inhibition from other effects on catalytic activity. Kinetic studies via in situ nuclear magnetic resonance spectroscopy indicated substantial product inhibition upon catalyst activity after several tens to several thousand turnovers, depending on specific conditions. Apparent product binding constants were obtained by fitting initial reaction rates at pH 7.0 and pH 10.5 to a Langmuir-Freundlich binding/adsorption model. For the fits, varying amounts/concentrations of candidate inhibitors were introduced before the start of catalytic hydrolysis. The derived binding constants proved suitable for quantitatively describing product inhibition effects upon reaction rates over the extended time course of simulant hydrolysis by aqua-ligand-bearing hexa-zirconium(IV) nodes.

The serious problem of pharmaceutical and personal care product pollution places great pressure on aquatic environments and human health. Herein, a novel coating photocatalyst was synthesized by adhering Ag-AgCl/WO 3 /g-C 3 N 4 (AWC) nanoparticles on a polydopamine (PDA)-modified melamine sponge (MS) through a facile layer-by-layer assembly method to degrade trimethoprim (TMP). The formed PDA coating was used for the anchoring of nanoparticles, photothermal conversion, and hydrophilic modification. TMP (99.9%; 4 mg/L) was removed in 90 min by the photocatalyst coating (AWC/PDA/MS) under visible light via a synergistic photocatalytic-photothermal performance route. The stability and reusability of the AWC/PDA/MS have been proved by cyclic experiments, in which the removal efficiency of TMP was still more than 90% after five consecutive cycles with a very little mass loss. Quantitative structure-activity relationship analysis revealed that the ecotoxicities of the generated intermediates were lower than those of TMP. Furthermore, the solution matrix effects on the photocatalytic removal efficiency were investigated, and the results revealed that the AWC/PDA/MS still maintained excellent photocatalytic degradation efficiency in several actual water and simulated water matrices. This work develops recyclable photocatalysts for the potential application in the field of water remediation.

New chitosan Schiff base (3EtO-4OH/Chit) and its 3EtO-4OH/Chit/Fe 2 O 3 nanocomposite were synthesized and characterized by FTIR, 1 H NMR, XRD, TGA, DSC and SEM. The result confirmed the preparation of 3EtO-4OH/Chit and its 3EtO-4OH/Chit/Fe 2 O 3 nanocomposite. The efficiency of the prepared catalysts was studied for the methyl orange (MO) removal from aqueous solution. The effect of adsorbent dose and contact time on the removal of dye has been studied. Their antibacterial activities were considered against two Gram positive (S. aureus and B. cereus) and two Gram negative (E. coli and P. aeruginosa) bacteria and the results showed that the activity of the 3EtO-4OH/Chit/Fe 2 O 3 is excellent and is more than chitosan and 3EtO-4OH/Chit. Thermogravimetry studies shows that the weight loss stages and the residual value at 600 °C are different for the two compounds. DSC curve of the title compounds 3EtO-4OH/Chit and 3EtO-4OH/Chit/Fe 2 O 3 is different from each other. The reason for this difference could be due to the presence of iron oxide nanoparticles in 3EtO-4OH/Chit/Fe 2 O 3 ., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

O -Dealkylation of the tyrosine kinase inhibitor lapatinib by cytochrome P450 3A enzymes is implicated in the development of lapatinib-induced hepatotoxicity. Conjugative metabolism of debenzylated lapatinib (M1) via glucuronidation and sulfation is thought to be a major detoxication pathway for lapatinib in preclinical species (rat and dog), limiting formation of the quinoneimine reactive metabolite. Glucuronidation of M1 by human recombinant UDP-glucuronosyltransferases (UGTs) has been reported in vitro; however, the relative UGT enzyme contributions are unknown, and the interspecies differences in the conjugation versus bioactivation pathways of M1 have not been fully elucidated. In the present study, reaction phenotyping experiments using human recombinant UGT enzymes and enzyme-selective chemical inhibitors demonstrated that UGT1A1 was the major hepatic UGT enzyme involved in lapatinib M1 glucuronidation. Formation of the M1-glucuronide by human liver microsomes from UGT1A1 -genotyped donors was significantly correlated with UGT1A1 activity as measured by 17 β -estradiol 3-glucuronidation ( R 2 = 0.90). Interspecies differences were found in the biotransformation of M1 in human, rat, and dog liver microsomal and 9000 g supernatant (S9) fractions via glucuronidation, sulfation, aldehyde oxidase-mediated oxidation, and bioactivation to the quinoneimine trapped as a glutathione (GSH) conjugate. Moreover, we demonstrated the sequential metabolism of lapatinib in primary human hepatocytes to the M1-glucuronide, M1-sulfate, and quinoneimine-GSH conjugate. M1 glucuronidation was highly correlated with the rates of M1 formation, suggesting that O -dealkylation may be the rate-limiting step in lapatinib biotransformation. Interindividual variability in the formation and clearance pathways of lapatinib M1 likely influences the hepatic exposure to reactive metabolites and may affect the risk for hepatotoxicity. SIGNIFICANCE STATEMENT: We used an integrated approach to examine the interindividual and interspecies differences in detoxication versus bioactivation pathways of lapatinib, which is associated with idiosyncratic hepatotoxicity. In addition to cytochrome P450 (P450)-mediated bioactivation, we report that multiple non-P450 pathways are involved in the biotransformation of the primary phenolic metabolite of lapatinib in vitro, including glucuronidation, sulfation, and aldehyde oxidase mediated oxidation. UGT1A1 was identified as the major hepatic enzyme involved in debenzylated lapatinib glucuronidation, which may limit hepatic exposure to the potentially toxic quinoneimine., Competing Interests: Financial Disclosure: The authors have no financial conflicts of interest to disclose., (Copyright © 2021 by The American Society for Pharmacology and Experimental Therapeutics.)

The present work pronounces the three phase partitioning (TPP)-facilitated preparation of porous cross-linked Candida antarctica lipase B (CaLB) aggregates (pCLEAs) for 5-Hydroxymethylfurfural (HMF) esters synthesis. CLEAs and pCLEAs of CaLB were prepared with eupergit as the support under the optimized conditions of pH 8.0, eupergit/protein ratio of 3.0:1.0, 50 mM cross-linker concentration and 3.3 mg/mL BSA concentration in 4 h. The optimum starch concentration for pCLEAs was 0.20%, m/v. The maximum biocatalytic load was 650 U/g (CLEAs) and 721 U/g (pCLEAs), and the immobilized biocatalysts were stable over a pH range of 6.0-9.0 and temperature range of (40-60)°C. The BET surface area of CLEAs and pCLEAs were 21.3 and 29.1 m 2 /g, respectively, and the catalytic efficiency of pCLEAs was 2.2-fold higher than that of CLEAs. Subsequently, the pCLEAs of CaLB were utilized for the manufacturing of industrially significant HMF esters. Under the optimized transesterification conditions, HMF conversion with pCLEAs CaLB was 1.41- and 1.25-fold higher than with free and CLEAs CaLB, respectively. The pCLEAs were reused upto 8 consecutive transesterification cycles and the produced HMF esters reduced the surface tension of water from 72 mN/m to 32.6 mN/m, proving its potential application as surface-active compounds., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Precise monitoring of the enzyme activity by a suitable modulator is one of the very fundamental aspects of drug designing that provides the opportunity to overcome the challenges of several diseases. Herein, inhibition of human Topoisomerase IIα enzyme which serves as a potential target site for several anti-cancer drugs is demonstrated by using ultra-small size gold nanoclusters (Au NCs) with the dimension comparable with size of the active site of the enzyme. Molecular dynamics simulation results demonstrate that the Au NCs strongly interact with the human Topo IIα enzyme at its active site or allosteric site depending on forms of enzyme. Additionally, binding energy and interaction profile provides the molecular basis of understanding of interactions of ultra-small size Au NCs and human Topo IIα enzyme. Enthalpy change (ΔH) and binding constant (K) are measured based on a sequential binding model of the Au NCs with the enzyme as demonstrated by the ITC study. Moreover, the in-vitro inhibition study of the catalytic activity of the enzyme and gel electrophoresis indicates that the ultra-small size Au NCs may be used as a potent inhibitor of human Topo IIα enzyme., (Copyright © 2020. Published by Elsevier B.V.)

Poly-(ADP-ribose) polymerase 1 and 2 (PARP1 and PARP2) are key enzymes in the DNA damage response. Four different inhibitors (PARPi) are currently in the clinic for treatment of ovarian and breast cancer. Recently, histone PARylation Factor 1 (HPF1) has been shown to play an essential role in the PARP1- and PARP2-dependent poly-(ADP-ribosylation) (PARylation) of histones, by forming a complex with both enzymes and altering their catalytic properties. Given the proximity of HPF1 to the inhibitor binding site both PARPs, we hypothesized that HPF1 may modulate the affinity of inhibitors toward PARP1 and/or PARP2. Here we demonstrate that HPF1 significantly increases the affinity for a PARP1 - DNA complex of some PARPi (i.e., olaparib), but not others (i.e., veliparib). This effect of HPF1 on the binding affinity of Olaparib also holds true for the more physiologically relevant PARP1 - nucleosome complex but does not extend to PARP2. Our results have important implications for the interpretation of PARP inhibition by current PARPi as well as for the design and analysis of the next generation of clinically relevant PARP inhibitors.

In biosynthesis of the pancreatic cancer drug streptozotocin, the tridomain nonheme-iron oxygenase SznF hydroxylates N δ and N ω ' of N ω -methyl-l-arginine before oxidatively rearranging the triply modified guanidine to the N -methyl- N -nitrosourea pharmacophore. A previously published structure visualized the monoiron cofactor in the enzyme's C-terminal cupin domain, which promotes the final rearrangement, but exhibited disorder and minimal metal occupancy in the site of the proposed diiron cofactor in the N- hydroxylating heme-oxygenase-like (HO-like) central domain. We leveraged our recent observation that the N -oxygenating µ-peroxodiiron(III/III) intermediate can form in the HO-like domain after the apo protein self-assembles its diiron(II/II) cofactor to solve structures of SznF with both of its iron cofactors bound. These structures of a biochemically validated member of the emerging heme-oxygenase-like diiron oxidase and oxygenase (HDO) superfamily with intact diiron cofactor reveal both the large-scale conformational change required to assemble the O 2 -reactive Fe 2 (II/II) complex and the structural basis for cofactor instability-a trait shared by the other validated HDOs. During cofactor (dis)assembly, a ligand-harboring core helix dynamically (un)folds. The diiron cofactor also coordinates an unanticipated Glu ligand contributed by an auxiliary helix implicated in substrate binding by docking and molecular dynamics simulations. The additional carboxylate ligand is conserved in another N -oxygenating HDO but not in two HDOs that cleave carbon-hydrogen and carbon-carbon bonds to install olefins. Among ∼9,600 sequences identified bioinformatically as members of the emerging HDO superfamily, ∼25% conserve this additional carboxylate residue and are thus tentatively assigned as N -oxygenases., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

The current communication reports the inhibitory effect of para-benzoquinone (p-BQ) on the structure and function of bovine liver catalase (BLC), a vital antioxidant enzyme. Both BLC and p-BQ were dissolved in respective buffers and the biophysical interaction was studied at physiological concentrations. For the first time our data reveals an enthalpy-driven interaction between BLC and p-BQ which is due to hydrogen bonding and van der Waals interactions. The binding affinity of p-BQ with BLC is nearly 2.5 folds stronger in MOPS buffer than Phosphate buffer. Importantly, the binding affinity between BLC and p-BQ was weak in HEPES buffer as compared to other buffers being the strongest in Tris buffer. Molecular docking studies reveal that binding affinity of p-BQ with BLC differ depending upon the nature of buffers rather than on the participating amino acid residues of BLC. This is further supported by the differential changes in secondary structures of BLC. The p-BQ-induced conformational change in BLC was evident from the reduced BLC activity in presence of different buffers in the following order, Phosphate>MOPS>Tris>HEPES. The absorbance peak of BLC was gradually increased and fluorescence spectra of BLC were drastically decreased when BLC to p-BQ molar ratio was incrementally enhanced from 0 to 10,000 times in presence of all buffers. Nevertheless, the declined activity of BLC was positively correlated with the reduced fluorescence and negatively correlated with the enhanced absorbance. Electrochemical study with cyclic voltammeter also suggests a direct binding of p-BQ with BLC in presence of different buffers. Thus, p-BQ-mediated altered secondary structure in BLC results into compromised activity of BLC., Competing Interests: Declaration of competing interest Authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Traumatic brain injury (TBI) is a sudden injury to the brain, accompanied by the production of large amounts of reactive oxygen and nitrogen species (RONS) and acute neuroinflammation responses. Although traditional pharmacotherapy can effectively decrease the immune response of neuron cells via scavenging free radicals, it always involves in short reaction time as well as rigorous clinical trial. Therefore, a noninvasive topical treatment method that effectively eliminates free radicals still needs further investigation. Methods: In this study, a type of catalytic patch based on nanozymes with the excellent multienzyme-like activity is designed for noninvasive treatment of TBI. The enzyme-like activity, free radical scavenging ability and therapeutic efficacy of the designed catalytic patch were assessed in vitro and in vivo . The structural composition was characterized by the X-ray diffraction, X-ray photoelectron spectroscopy and high-resolution transmission electron microscopy technology. Results: Herein, the prepared Cr-doped CeO 2 (Cr/CeO 2 ) nanozyme increases the reduced Ce 3+ states, resulting in its enzyme-like activity 3-5 times higher than undoped CeO 2 . Furthermore, Cr/CeO 2 nanozyme can improve the survival rate of LPS induced neuron cells via decreasing excessive RONS. The in vivo experiments show the Cr/CeO 2 nanozyme can promote wound healing and reduce neuroinflammation of mice following brain trauma. The catalytic patch based on nanozyme provides a noninvasive topical treatment route for TBI as well as other traumas diseases. Conclusions: The catalytic patch based on nanozyme provides a noninvasive topical treatment route for TBI as well as other traumas diseases., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Full degradation of acetaminophen (paracetamol) in aqueous solution was investigated at room temperature through heterogeneous iron nano-structured as catalyst in this article. Iron Nano-structured was prepared through simple hydrothermal processes using Iron oxide (Fe 2 O 3 ) as precursor. The catalytic activity of as prepared Nano-catalyst (NC) was investigated in the degradation of the acetaminophen as an environmental pollutant, commonly called paracetamol, under different operating parameters like pH, dosages of acetaminophen and dose of NC. Remarkable differences in IR spectra were observed after reaction which showed complete degradation of 15 ppm of Acetaminophen using 0.1 g of nano-structured with the recovery of NC followed by its activity four times with full catalytic performance.

Metastasis and spread are currently the main factors leading to high mortality of cancer, so developing a synergetic antitumor strategy with high specificity and hypotoxicity is in urgent demand. Based on the design concept of "nanocatalytic medicine", multifunctional nanotherapeutic agent FePt@COP-FA nanocomposites (FPCF NCs) are developed for cancer treatment. Specifically, in the tumor microenvironment (TME), FePt could catalyze intracellular over-expressed H2O2 to generate highly active hydroxyl radicals (˙OH), which could not only induce the apoptosis of tumor cells, but also activate the "ferroptosis" pathway resulting in the lipid peroxide accumulation and ferroptotic cell death. Moreover, owing to the excellent photothermal effect, the FPCF NCs could effectively ablate primary tumors under near-infrared (NIR) laser irradiation and produce numerous tumor-associated antigens in situ. With the assistance of a checkpoint blockade inhibitor, anti-CTLA4 antibody, the body's specific immune response would be initiated to inhibit the growth of metastatic tumors. In particular, such synergistic therapeutics could produce an effective immunological memory effect, which could prevent tumor metastasis and recurrence again. In summary, the FPCF NC is an effective multifunctional antitumor therapeutic agent for nanocatalytic/photothermal/checkpoint blockade combination therapy, which exhibits great potential in nanocatalytic anticancer therapeutic applications.

Present study focuses on the organic solvent and ionic liquid compatibility of lignolytic enzymes produced from H. aswanensis strain ABC_IITR. Lignin peroxidase (LiP), Manganese Peroxidase (MnP) and Laccase (Lac) obtained by Solid State Fermentation (SSF) of wheat bran. In this work, lignolytic enzymes subjected to 20-40% (v/v) organic solvents, metals ions, and cholinium laurate based ionic liquid (CLIL). Use of 40% (v/v) of pyridine along with 1.5 M NaCl and 0.15 mM CLIL in reaction system increased the bio-catalytic activity of lignolytic enzymes whereas metal ions like Fe increased LiP and MnP activities, and Cu enhanced laccase activity compared to control. The inherent stability of lignolytic enzymes in CLIL was not affected significantly whereas it decreased in pyridine reaction system. Further, in Kalson lignin degradation study, higher degradation achieved in CLIL (generate less saline waste) as compared to 40% (v/v) pyridine reaction system. In MnP catalyzed system, use of Glutathione (GSH) as mediator had resulted in maximum reduction of lignin weight of 40.84 and 31.83% in 40% (v/v) pyridine (1.5 NaCl) and 0.15 mM CLIL, respectively. This is the first report on lignolytic enzymes of haloarchaea with capability to get activated in organic solvent and cholinium laurate based ionic liquid., (Copyright © 2020. Published by Elsevier B.V.)

Under mild acidic conditions, various metal derivatives of tetrakis(4- N -methylpyridinium)porphyrin (gold(III), AuT 4 ; cobalt(III), CoT 4 ; manganese(III), MnT 4 and zinc(II), ZnT 4 ) catalytically promote the supramolecular assembling process of the diacid 5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrin (H 2 TPPS 4 ) into J-aggregates. The aggregation kinetics have been treated according to a well-established model that involves the initial formation of a critical nucleus containing m porphyrin units, followed by autocatalytic growth, in which the rate evolves as a power of time. An analysis of the extinction time traces allows to obtain the rate constants for the auto-catalyzed pathway, k c , and the number of porphyrins involved in the initial seeding. The aggregation kinetics have been investigated at fixed H 2 TPPS 4 concentration as a function of the added metal derivatives MT 4 . The derived rate constants, k c , obey a rate law that is first order in [MT 4 ] and depend on the specific nature of the catalyst in the order AuT 4 > CoT 4 > MnT 4 > ZnT 4 . Both resonance light scattering (RLS) intensity and extinction in the aggregated samples increase on increasing [MT 4 ]. With the exception of AuT 4 , the final aggregated samples obtained at the highest catalyst concentration exhibit a negative Cotton effect in the J-band region, evidencing the occurrence of spontaneous symmetry breaking. The role of the nature of the metal derivative in terms of overall charge and presence of axial groups will be discussed.

ZntA from Escherichia coli confers resistance to toxic concentrations of Pb 2+ , Zn 2+ , and Cd 2+ . It is a member of the P 1B -ATPase transporter superfamily, which includes the human Cu + -transporting proteins ATP7A and ATP7B. P 1B -type ATPases typically have a hydrophilic N-terminal metal-binding domain and eight transmembrane helices. A splice variant of ATP7B was reported, which has 100-fold higher night-specific expression in the pineal gland; it lacks the entire N-terminal domain and the first four transmembrane helices. Here, we report our findings with Δ231-ZntA, a similar truncation we created in ZntA. Δ231-ZntA has no in vivo and greatly reduced in vitro activity. It binds one metal ion per dimer at the transmembrane site, with a 15-19000-fold higher binding affinity, indicating highly significant changes in the dimer structure of Δ231-ZntA relative to that of ZntA. Cd 2+ has the highest affinity for Δ231-ZntA, in contrast to ZntA, which has the highest affinity for Pb 2+ . Site-specific mutagenesis of the metal-binding residues, 392 Cys, 394 Cys, and 714 Asp, showed that there is considerable flexibility at the metal-binding site, with any two of these three residues able to bind Zn 2+ and Pb 2+ unlike in ZntA. However, Cd 2+ binds to only 392 Cys and 714 Asp, with 394 Cys not involved in Cd 2+ binding. Three-dimensional homology models show that there is a dramatic difference between the ZntA and Δ231-ZntA dimer structures, which help to explain these observations. Therefore, the first four transmembrane helices in ZntA and P 1B -type ATPases play an important role in maintaining the correct dimer structure.

Immobilized lipase has become one of the most important biocatalytic systems used for many industrial purposes, especially in the fuel, pharmaceutical, cosmetics, and food industries. Immobilized lipase has advantages of good chemical and thermal stability, in addition to reuse and easy recovery, allowing it to serve as a replacement for chemical catalysts in industrial applications. In this regard, there has been rapidly growing interest in developing new support materials (SMs) and new protocols for lipase immobilization (LI). In this review, various types of SMs used for LI are extensively discussed, with clarification of their advantages and disadvantages. In addition, different methods of LI are explained with a particular emphasis on their respective benefits and drawbacks. The applications of immobilized lipase in producing biodiesel as well as other organic synthesis processes are summarized with a mention of future prospects and directions., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Current study deals with the comparative study related to immobilization of maltase using synthetic (polyacrylamide) and non-synthetic (calcium alginate, agar-agar and agarose) polymers via entrapment technique. Polyacrylamide beads were formed by cross-linking of monomers, agar-agar and agarose through solidification while alginate beads were prepared by simple gelation. Results showed that the efficiency of enzyme significantly improved after immobilization and among all tested supports agar-agar was found to be the most promising and biocompatible for maltase in terms of immobilization yield (82.77%). The catalytic behavior of maltase was slightly shifted in terms of reaction time (free enzyme, agarose and polyacrylamide: 5.0 min; agar-agar and alginate: 10.0 min), pH (free enzyme, alginate and polyacrylamide: 6.5; agar-agar, agarose: 7.0) and temperature (free enzyme: 45 °C; alginate: 50 °C; polyacrylamide: 55 °C; agarose: 60 °C; agar-agar: 65 °C). Stability profile of immobilized maltase also revealed that all the supports utilized have significantly enhanced the activity of maltase at higher temperatures then its free counterpart. However, recycling data showed that agar-agar entrapped maltase retained 20.0% of its initial activity even after 10 cycles followed by agarose (10.0%) while polyacrylamide and alginate showed no activity after 8 and 6 cycles respectively., Competing Interests: Declaration of competing interest This publication is approved by all authors and they do not have any conflict of interest regarding any financial, personal or other relationships with any other people or organizations., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Xylan and pectin are major structural components of plant cell walls. There are two independent catabolic pathways for xylan and pectin. UxaE bridges these two pathways by reversibly epimerizing D-fructuronate and D-tagaturonate. The crystal structure of UxaE from Cohnella laeviribosi (ClUxaE) shows a core scaffold of TIM-barrel with a position-changing divalent metal cofactor. ClUxaE has the flexible metal-coordination loop to allow the metal shift and the extra domains to bind a phosphate ion in the active site, which are important for catalysis and substrate specificity. Elucidation of the structure and mechanism of ClUxaE will assist in understanding the catalytic mechanism of UxaE family members, which are useful for processing both xylan and pectin-derived carbohydrates for practical and industrial purposes, including the transformation of agricultural wastes into numerous valuable products., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020. Published by Elsevier B.V.)

Polymerase eta (Polη) is a translesion synthesis DNA polymerase directly linked to cancer development. It can bypass several DNA lesions thereby rescuing DNA damage-stalled replication complexes. We previously presented evidence implicating Saccharomyces cerevisiae Polη in transcription elongation, and identified its specific RNA extension and translesion RNA synthetic activities. However, RNA synthesis by Polη proved rather inefficient under conditions optimal for DNA synthesis. Searching for factors that could enhance its RNA synthetic activity, we have identified the divalent cation of manganese. Here, we show that manganese triggers drastic changes in the activity of Polη. Kinetics experiments indicate that manganese increases the efficiency of ribonucleoside incorporation into RNA by ~400-2000-fold opposite undamaged DNA, and ~3000 and ~6000-fold opposite TT dimer and 8oxoG, respectively. Importantly, preference for the correct base is maintained with manganese during RNA synthesis. In contrast, activity is strongly impaired, and base discrimination is almost lost during DNA synthesis by Polη with manganese. Moreover, Polη shows strong preference for manganese during RNA synthesis even at a 25-fold excess magnesium concentration. Based on this, we suggest that a new regulatory mechanism, selective metal cofactor utilization, modulates the specificity of Polη helping it to perform distinct activities needed for its separate functions during replication and transcription.

An efficient and environmentally sustainable synthetic protocol has been presented to synthesize structurally diverse spiroxindoles spiroannulated with indenopyrroloimidazoles, pyranopyrroloimidazoles, chromenopyrroloimidazoles, and imidazopyrrolopyrimidines involving three-component reaction of isatins, hydantoin, and β-diketones in the presence of green and sustainable bio-organic catalyst, β-amino acid, 2-aminoethanesulfonic acid (taurine), in aqueous media. The synthetic efficiency, operational simplicity, and reusability of catalyst make the present synthetic protocol cost effective, time efficient, and eco-friendly to synthesize molecules with structural diversity and molecular complexity and expected to contribute significantly not only to drug discovery research but also to pharmaceutical and medicinal chemistry.

Three-component reaction between arylglyoxals, 1,3-dicetones and enaminoketones leads to new polyfunctionalized tetraone derivatives which may be easily converted to polyfunctionalized pyrroles. Reactions were conducted in water or water-ethanol mixture as green solvents, and all products were isolated by simple washing of the resulting solids with diethyl ether.

A simple preparation of catalysts with high catalytic activity and superior cycling stability is very desirable. In this contribution, magnetic carbon nanotube functionalized by polythiophene (CNT-Fe 3 O 4 -PTh) acts as an efficient and retrievable host for copper nanoparticles to prepare CNT-Fe 3 O 4 -PTh-Cu(I) as a nontoxic and inexpensive catalyst. FT-IR, TGA, EDX, VSM, XRD, FE-SEM, TEM, and AAS techniques were employed to characterize the structure of the synthesized magnetic heterogeneous nanocomposite. Thereafter, the catalytic application of the catalyst was evaluated for the phosphine- and palladium-free Suzuki-Miyaura cross-coupling reaction in water/ethanol as a green media in short reaction times with good to excellent yields. Various derivatives of biaryl compounds were synthesized by reaction of aryl halides and phenylboronic acid. Simple methodology and easy workup, short reaction times, elimination of volatile and toxic solvents, biocompatible reaction conditions, and high yields are some advantages of this protocol. Moreover, the catalyst showed a good reusability owing to its magnetic properties and was recycled several times without appreciable decrease in its catalytic efficiency. Copper(I) nanoparticles supported on magnetic carbon nanotube functionalized by polythiophene (CNT-Fe 3 O 4 -PTh-Cu(I)), was prepared and used for ligand- and palladiumfree Suzuki-Miyaura cross-coupling reaction in water/ethanol as a green media in short reaction times with good to excellent yields. Reusability and stability tests demonstrated that the as prepared catalyst can be recycled with a negligible loss of its activity.

2-Amino-3-cyano-4H-chromenes are structural core motifs that received increasing attention in the last years due to their interesting potential pharmacological properties. In this review, the synthetic methods for these compounds are classified based on the type of catalyst in the pertinent reactions. In addition, the wide range of pharmacological properties of these compounds is covered in a separate section.

A novel five-component diastereoselective synthesis of polysubstituted 2-piperidinones is reported. The Knoevenagel condensation-Michael addition-Mannich cascade of two equivalents of aromatic aldehydes, nitriles, dialkyl malonates and ammonium acetate or aqueous ammonia in alcohols provides convenient access to alkyl (3SR,4RS,6SR)-5,5-dicyano-2-oxo-4,6-diarylpiperidine-3-carboxylates with three stereocenters in 52-90% or dialkyl (2SR,3RS,4RS,5SR)-2,4-diaryl-3-cyano-6-oxopiperidine-3,5-dicarboxylates with four stereocenters in 38-88%. The formation of products was highly stereoselective, with only one diastereomer formed. Ammonium acetate or aqueous ammonia plays a role both as a catalyst and as a nitrogen source. 2,4,6-triaryl-3,3,5,5-tetracyanopiperidines were obtained as a side products in the reactions with nitro-substituted aldehydes or with ethyl and n-propyl cyanoacetates. A series of 14 2-piperidinones and piperidines was assessed for antimicrobial activity against a panel of five bacteria and two fungi; no significant activity was observed. Two side piperidines with nitro substituents in aromatic ring possess bacteriostatic action against S. aureus ATCC 43300 and A. baumannii ATCC 19606 at 32 ug/mL.

Use of cheaper and recyclable materials contributes positively to economic growth with environmental sustainability. We report the prospect of utilizing red brick clay as catalyst, which exhibited excellent activity in rapid one-pot four-component condensation of 1,2,4,5-tetrasubstituted imidazoles with high conversion and yields (91-96%) in aqueous medium at 60 °C in short reaction times (25-40 min). The red brick clay material was fully characterized by XRD, FT-IR, SEM, TEM, EDX and BET analyses. Red brick clay consisted of oxides of Si (20.38%), Fe (19.55%), Al (14.30%) and minor amounts of Ca (3.60%) and Mg (1.68%). The slate-like-shaped structure morphology and flaky appearance of inexpensive solid clay material proved competent material for the synthesis of 15 novel 1,2,4,5-tetrasubstituted imidazole derivatives. In addition, the advantages of the eco-friendly method are non-toxicity and re-usability of the catalyst. Reaction offers 78% atom economy and 84% carbon capture.

Inhibition of pancreatic lipase (PL) is considered one of the important therapeutic interventions against obesity. In the present study, the inhibition of porcine (mammalian) PL (PPL) by two tripeptides glutathione (GSH) and s-allyl glutathione (SAG) was studied. In vitro kinetic analysis was done to determine the inhibition of GSH and SAG against PPL. The binding of GSH and SAG with PPL was elucidated by fluorescence spectroscopy analysis. Docking and molecular dynamics (MD) simulation analysis was carried out to understand the intermolecular interaction between both GSH and SAG with PPL as well as human PL (HPL). Both GSH and SAG inhibited PPL in mixed non-competitive manner. The IC 50 value for GSH and SAG against PPL was found to be 2.97 and 6.4 mM, respectively. Both GSH and SAG quenched the intrinsic fluorescence of PPL through static quenching that is through forming complex with the PPL. SAG and GSH interacted with amino acids involved in catalysis of both PPL and HPL. MD simulation showed interactions of SAG and GSH with both PPL and HPL were stable. These results would lead to the further studies and application of GSH and SAG against obesity through inhibition of PL., (Copyright © 2020 Elsevier B.V. All rights reserved.)

DNA topoisomerase II (topo II) is an essential enzyme that regulates DNA topology by DNA cleavage and re-ligation. In vertebrates, there are two isozymes, α and β. The C-terminal domain (CTD) of the isozymes, which shows a low degree of sequence homology between α and β, is involved in each isozyme-specific intracellular behavior. The CTD of topo IIβ is supposedly involved in topo II regulation. Topo IIβ is maintained in an inactive state in the nucleoli by the binding of RNA to the 50-residue region termed C-terminal regulatory domain (CRD) present in the CTD. Although in vitro biochemical analysis indicates that the CTD of topo IIβ has DNA binding activity, it is unclear whether CTD influences catalytic reaction in the nucleoplasm. Here, we show that the proximal CTD (hereafter referred to as pCTD) of rat topo IIβ, including the CRD, is involved in the catalytic reaction in the nucleoplasm. We identified the pCTD as a domain with DNA binding activity by in vitro catenation assay and electrophoretic mobility shift assay. Fluorescence recovery after photo-bleaching (FRAP) analysis of pCTD-lacking mutant (ΔpCTD) showed higher mobility in nucleoplasm than that of the wild-type enzyme, indicating that the pCTD also affected the nuclear dynamics of topo IIβ. ICRF-193, one of the topo II catalytic inhibitors, induces the formation of closed-clamp intermediates of topo II. Treatment of ΔpCTD with ICRF-193 significantly decreased the efficiency of closed-clamp formation. Altogether, our data indicate that the binding of topo IIβ to DNA through the pCTD is required for the catalytic reaction in the nucleoplasm., Competing Interests: The authors have declared that no competing interests exist.

Boronic acids have been successfully employed as inhibitors of hydrolytic enzymes. Typically, an enzymatic nucleophile catalyzing hydrolysis adds to the electrophilic boron atom forming a tetrahedral species that mimics the intermediate(s)/transition state(s) for the hydrolysis reaction. We show that para -substituted phenylboronic acids (PBAs) are potent competitive inhibitors of mandelate racemase (MR), an enzyme that catalyzes a 1,1-proton transfer rather than a hydrolysis reaction. The K i value for PBA was 1.8 ± 0.1 μM, and p -Cl-PBA exhibited the most potent inhibition ( K i = 81 ± 4 nM), exceeding the binding affinity of the substrate by ∼4 orders of magnitude. Isothermal titration calorimetric studies with the wild-type, K166M, and H297N MR variants indicated that, of the two Brønsted acid-base catalysts Lys 166 and His 297, the former made the greater contribution to inhibitor binding. The X-ray crystal structure of the MR·PBA complex revealed the presence of multiple H-bonds between the boronic acid hydroxyl groups and the side chains of active site residues, as well as formation of a His 297 N ε2 -B dative bond. The dramatic upfield change in chemical shift of 27.2 ppm in the solution-phase 11 B nuclear magnetic resonance spectrum accompanying binding of PBA by MR was consistent with an sp 3 -hybridized boron, which was also supported by density-functional theory calculations. These unprecedented findings suggest that, beyond substituting boron at carbon centers participating in hydrolysis reactions, substitution of boron at the acidic carbon center of a substrate furnishes a new approach for generating inhibitors of enzymes catalyzing the deprotonation of carbon acid substrates.

Tyrosine biosynthesis via the shikimate pathway is absent in humans and other animals, making it an attractive target for next-generation antibiotics, which is increasingly important due to the looming proliferation of multidrug-resistant pathogens. Tyrosine biosynthesis is also of commercial importance for the environmentally friendly production of numerous compounds, such as pharmaceuticals, opioids, aromatic polymers, and petrochemical aromatics. Prephenate dehydrogenase (PDH) catalyzes the penultimate step of tyrosine biosynthesis in bacteria: the oxidative decarboxylation of prephenate to 4-hydroxyphenylpyruvate. The majority of PDHs are competitively inhibited by tyrosine and consist of a nucleotide-binding domain and a dimerization domain. Certain PDHs, including several from pathogens on the World Health Organization priority list of antibiotic-resistant bacteria, possess an additional ACT domain. However, biochemical and structural knowledge was lacking for these enzymes. In this study, we successfully established a recombinant protein expression system for PDH from Bacillus anthracis (BaPDH), the causative agent of anthrax, and determined the structure of a BaPDH ternary complex with NAD + and tyrosine, a binary complex with tyrosine, and a structure of an isolated ACT domain dimer. We also conducted detailed kinetic and biophysical analyses of the enzyme. We show that BaPDH is allosterically regulated by tyrosine binding to the ACT domains, resulting in an asymmetric conformation of the BaDPH dimer that sterically prevents prephenate binding to either active site. The presented mode of allosteric inhibition is unique compared to both the competitive inhibition established for other PDHs and to the allosteric mechanisms for other ACT-containing enzymes. This study provides new structural and mechanistic insights that advance our understanding of tyrosine biosynthesis in bacteria. ENZYMES: Prephenate dehydrogenase from Bacillus anthracis (PDH): EC database ID: 1.3.1.12. DATABASES: Coordinates and structure factors have been deposited in the Protein Data Bank (PDB) with accession numbers PDB ID: 6U60 (BaPDH complex with NAD + and tyrosine), PDB ID: 5UYY (BaPDH complex with tyrosine), and PDB ID: 5V0S (BaPDH isolated ACT domain dimer). The diffraction images are available at http://proteindiffraction.org with DOIs: https://doi.org/10.18430/M35USC, https://doi.org/10.18430/M35UYY, and https://doi.org/10.18430/M35V0S., (© 2019 Federation of European Biochemical Societies.)

Phosphodiesterase type 2A (PDE2A) has received considerable interest as a molecular target for treating central nervous system diseases that affect memory, learning, and cognition. In this paper, the authors present the discovery of small molecules that have a novel modality of PDE2A inhibition. PDE2A possesses GAF-A and GAF-B domains and is a dual-substrate enzyme capable of hydrolyzing both cGMP and cAMP, and activation occurs through cGMP binding to the GAF-B domain. Thus, positive feedback of the catalytic activity to hydrolyze cyclic nucleotides occurs in the presence of appropriate concentrations of cGMP, which binds to the GAF-B domain, resulting in a "brake" that attenuates downstream cyclic nucleotide signaling. Here, we studied the inhibitory effects of some previously reported PDE2A inhibitors, all of which showed impaired inhibitory effects at a lower concentration of cGMP (70 nM) than a concentration effective for the positive feedback (4 μM). This impairment depended on the presence of the GAF domains but was not attributed to binding of the inhibitors to these domains. Notably, we identified PDE2A inhibitors that did not exhibit this behavior; that is, the inhibitory effects of these inhibitors were as strong at the lower concentration of cGMP (70 nM) as they were at the higher concentration (4 μM). This suggests that such inhibitors are likely to be more effective than previously reported PDE2A inhibitors in tissues of patients with lower cGMP concentrations.

A series of nine new 2,3-disubstituted 4(3H)-quinazolin-4-one derivatives was furnished starting from the 2-propyl-4(3H)-quinazo-line-4-one (2). The reinvestigation of the key starting quinazolinone 2 was performed under microwave irradiation (MW) and solvent-free conditions. Combination of MW and phase-transfer catalysis using tetrabutylammonium benzoate (TBAB) as a novel neutral ionic catalyst was used for carrying out N-alkylation and condensation reactions of compound 2 as a simple, efficient and eco-friendly technique. The structure of the synthesized compounds was elucidated using different spectral and chemical analyses. In vitro antimicrobial activity of the compounds was investigated against four bacterial and two fungal strains; very modest activity was achieved. Some of the synthesized compounds were screened for their antitumor activity against different human tumor cell lines. The screened compounds exhibited a significant antitumor activity on some of the cancer cell lines, melanoma (SK-MEL-2), ovarian cancer (IGROV1), renal cancer (TK-10), prostate cancer (PC-3), breast cancer (MCF7) and colon cancer (HT29). The most active, even more active than the reference 5-fluorouracil, were found to be ethyl 4-[(4-oxo-2-propylquinazolin-3(4H)-yl)methyl]benzoate (3c), 3-{2-[6-(pyrrolidin-1-yl-sulfonyl)-1,2,3,4-tetrahydroquinoline]-2-oxoethyl}-2-propylquinazolin--4(3H)-one (3e), N'-[(E)-(2H-1,3-benzodioxo-5-yl)methylidene]-2-(4-oxo-2-propylquinazolin-3(4H)-yl)acetohydrazide (10a), N'-[(E)-(4-hydroxyphenyl)methylidene]-2-(4-oxo-2-propylquinazo-lin-3(4H) -yl)acetohydrazide (10b) and N'-[(E)-(4-nitrophenyl)methyl idene]-2-(4-oxo-2-propylquinazolin-3(4H)-yl)acetohydrazide (10c).

Mo-Fe catalysts with different Mo dispersions were synthesized with fast (Cat-FS, 600 r·min -1 ) or slow stirring speed (Cat-SS, 30 r·min -1 ) by the coprecipitation method. Improving the stirring speed strengthened the mixing of the solution and increased the dispersion of particles in the catalyst, which exhibited favorable activity and selectivity. The byproduct (dimethyl ether (DME)) selectivity increased from 2.3% to 2.8% with Cat-SS, while it remained unchanged with Cat-FS in a stability test. The aggregation of particles and thin Mo-enriched surface layer decreased the catalyst surface area and slowed down the reoxidation of reduced active sites with Cat-SS, leaving more oxygen vacancies which promoted the formation of DME by the nonoxidative channel.

The pyridinium oximes are known esterolytic agents, usually classified in the literature as catalysts, which mimic the catalytic mode of hydrolases. Herein, we combined kinetic and computational studies of the pyridinium-4-oxime-mediated acetylthiocholine (AcSCh + ) hydrolysis to provide novel insights into their potential catalytic activity. The N -methyl- and N -benzylpyridinium-4-oximes have been tested as oximolytic agents toward the AcSCh + , while the newly synthesized O -acetyl- N -methylpyridinium-4-oxime iodide was employed for studying the consecutive hydrolytic reaction. The relevance of the AcSCh + hydrolysis as a competitive reaction to AcSCh + oximolysis was also investigated. The reactions were independently studied spectrophotometrically and rate constants, k oxime , k w and k OH , were evaluated over a convenient pH-range at I = 0.1 M and 25 °C. The catalytic action of pyridinium-4-oximes comprises two successive stages, acetylation (oximolysis) and deacetylation stage (pyridinium-4-oxime-ester hydrolysis), the latter being crucial for understanding the whole catalytic cycle. The complete mechanism is presented by the free energy reaction profiles obtained with (CPCM)/M06-2X/6-311++G(2df,2pd)//(CPCM)/M06-2X/6-31+G(d) computational model. The comparison of the observed rates of AcSCh + oximolytic cleavage and both competitive AcSCh + and consecutive pyridinium-4-oxime-ester hydrolytic cleavage revealed that the pyridinium-4-oximes cannot be classified as non-enzyme catalyst of the AcSCh + hydrolysis but as the very effective esterolytic agents.

Porous Pt electrocatalysts have been developed as an example of carbon-free porous metal catalysts in anticipation of polymer electrolyte membrane (PEM) fuel cells and PEM water electrolyzers through the assembly of the metal precursor and surfactant. In this study, porous Pt was structurally evaluated and found to have a porous structure composed of connected Pt particles. The resulting specific electrochemical surface area (ECSA) of porous Pt was 12.4 m 2 g -1 , which was higher than that of commercially available Pt black. Accordingly, porous Pt showed higher oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) activity than Pt black. When the activity was compared to that of a common carbon-supported electrocatalyst, Pt/ketjen black (KB), porous Pt showed a comparable ORR current density (2.5 mA cm -2 at 0.9 V for Pt/KB and 2.1 mA cm -2 at 0.9 V for porous Pt), and OER current density (6.8 mA cm -2 at 1.8 V for Pt/KB and 7.0 mA cm -1 at 1.8 V), even though the ECSA of porous Pt was only one-sixth that of Pt/KB. Moreover, it exhibited a higher durability against 1.8 V. In addition, when catalyst layers were spray-printed on the Nafion ® membrane, porous Pt displayed more uniform layers in comparison to Pt black, showing an advantage in its usage as a thin layer.

Inhibitors targeting the general RNA polymerase II (RNAPII) transcription machinery are candidate therapeutics in cancer and other complex diseases. Here, we review the molecular targets and mechanisms of action of these compounds, framing them within the steps of RNAPII transcription. We discuss the effects of transcription inhibitors in vitro and in cellular models (with an emphasis on cancer), as well as their efficacy in preclinical and clinical studies. We also discuss the rationale for inhibiting broadly acting transcriptional regulators or RNAPII itself in complex diseases.

Poly(ADP-ribose) polymerase-1 (PARP-1) is an abundant and ubiquitous nuclear enzyme that catalyzes the transfer of ADP-ribose from donor NAD + molecules to specific amino acids on substrate proteins. The catalytic activity of PARP-1 has long been known to be allosterically stimulated by the free ends of DNA, such as those found at double-strand breaks in the genome. A number of studies have also shown that the catalytic activity of PARP-1 can also be stimulated by various types of RNA. A recent study by Nakamoto et al., however, has contradicted these results, concluding that the apparent stimulatory activity of the RNAs was due to contaminating DNA in the RNA preparations used in the biochemical assays. Here we show using a carefully controlled set of biochemical assays that DNA-free, in vitro-transcribed, PARP-1-interacting snoRNAs can stimulate PARP-1 catalytic activity. We confirmed the activation of PARP-1 by snoRNAs using a chemically synthesized snoRNA, as well as CRISPR/Cas9-mediated knockout of snoRNAs in cells. Finally, we provide a set of considerations and experimental conditions for the careful evaluation of RNA-stimulated PARP-1 catalytic activity that will help researchers avoid artifacts.

Activation of the mitogen-activated protein kinase (MAPK) signaling pathway regulated by human MAP kinase 1 (MEK1) is associated with the carcinogenesis and progression of numerous cancers. In addition, two active mutations (P124S and E203K) have been reported to enhance the activity of MEK1, thereby eventually leading to the tumorigenesis of cancer. Trametinib is an MEK1 inhibitor for treating EML4-ALK-positive, EGFR-activated, and KRAS-mutant lung cancers. Therefore, in this study, molecular docking and molecular dynamic (MD) simulations were performed to explore the effects of inactive/active mutations (A52V/P124S and E203K) on the conformational changes of MEK1 and the changes in the interaction of MEK1 with trametinib. Moreover, steered molecular dynamic (SMD) simulations were further utilized to compare the dissociation processes of trametinib from the wild-type (WT) MEK1 and two active mutants (P124S and E203K). As a result, trametinib had stronger interactions with the non-active MEK1 (WT and A52V mutant) than the two active mutants (P124S and E203K). Moreover, two active mutants may make the allosteric channel of MEK1 wider and shorter than that of the non-active types (WT and A52V mutant). Hence, trametinib could dissociate from the active mutants (P124S and E203K) more easily compared with the WT MEK1. In summary, our theoretical results demonstrated that the active mutations may attenuate the inhibitory effects of MEK inhibitor (trametinib) on MEK1, which could be crucial clues for future anti-cancer treatment.

Penicillin V is a bacteriolytic β-lactam antibiotic drug. In the present work, we investigated the inhibitory effect of Penicillin V on the activity of mushroom tyrosinase for the first time. The molecular mechanism for the inhibition of tyrosinase by Penicillin V was investigated by means of kinetics analysis, fluorescence quenching and molecular docking techniques. The results showed that Penicillin V could inhibit both monophenolase and diphenolase activities with IC 50 of 16.6 ± 0.5 and 11.0 ± 0.2 mmol/L, respectively. The inhibitory type of Penicillin V on mushroom was mixed type, and the values of K I and K IS were 13.46 and 17.26 mmol/L, respectively. The fluorescence quenching and molecular docking showed that Penicillin V could form static interaction near the catalytic pocket of the enzyme to hinder the transportation of substrate to the active site, as well as reduce the copper plasticity for catalysis. Our results contributed to the usage of Penicillin V as a novel tyrosinase inhibitor with dual effect in field of antimicrobial and food preservation and could also provide guidance for the design of novel tyrosinase inhibitors.

The class III histone deacetylase sirtuin 6 (SIRT6) modulates numerous functions in the cell by deacetylating histone lysine residues. Interestingly, SIRT6's efficiency in in vitro experiments is far greater against substrates carrying long-chain fatty acyl modifications such as myristoylated lysine compared with acetylated counterparts, but the deacetylase activity can be stimulated by fatty acids and small-molecule allosteric modulators. A new study helps to explain this puzzling activation using a novel activator, thorough kinetic investigation, and mutagenesis studies. These data help elucidate the molecular requirements for activation of SIRT6 and provide a foundation for development of activators for therapeutic purposes., (© 2020 Nielsen and Olsen.)

Nanomaterials with enzyme-like activity have attracted much attention recently. Herein, we report the synthesis of a new type of 2D MXene-Ti 3 C 2 /CuS nanocomposites with peroxidase-like activity using a simple hydrothermal approach. Significantly, compared with the individual MXene-Ti 3 C 2 nanosheets or CuS nanoparticles, the MXene-Ti 3 C 2 /CuS nanocomposites show a synergistically enhanced peroxidase-like activity and can be used as an efficient mimetic peroxidase to catalyze the reaction of 3,3,5,5-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide (H 2 O 2 ), causing a blue color change. Kinetic studies reveal that the MXene-Ti 3 C 2 /CuS nanocomposites have a higher catalytic activity to TMB than their single components, and the catalytic reaction follows the ping-pong mechanism. The MXene-Ti 3 C 2 /CuS nanocomposites are used for the colorimetric determination of cholesterol with a linear range of 10-100 μM and a limit of detection (LOD) of 1.9 μM. Our results show that the MXene-Ti 3 C 2 /CuS nanocomposites based colorimetric cholesterol biosensor is cost-effective, sensitive, and selective, which has potential application in H 2 O 2 and cholesterol detection and clinic medicine diagnostics., (Copyright © 2019 Elsevier B.V. All rights reserved.)