Tailoring a robust nanozyme formulation based on surfactant stabilized lipase immobilized onto newly fabricated magnetic silica anchored graphene nanocomposite: Aggrandized stability and application.

Candida rugosa lipase (CRL) was treated with surfactants and immobilized onto the novel formulated magnetic graphene anchored silica nanocomposite (Fe3O4/SiO2/Gr NC). For this purpose, the surface of lipase was initially coated with Triton-X 100 and cetyltrimethylammonium bromide surfactants, to stabilize enzyme in its open form and was then adsorbed onto aminated Fe3O4/SiO2/Gr NC. Glutaraldehyde (GA) was then utilized to cross-link the adsorbed lipase onto the NC. The fabricated NC and conjugated lipase was characterized by various techniques such as FT-IR, XRD, TGA, SEM, TEM, CLSM, CD and Fluorescence spectroscopy. The magnetic character of the as-synthesized NC was verified by AGM investigation. CD and fluorescence spectroscopic analysis demonstrated slight structural rearrangements in lipase upon conjugation. The surfactant stabilized immobilized lipase demonstrated significantly enhanced thermostability, tolerance to various metal ions and inhibitors. The immobilization yield obtained owing to lipase interfacial activation by Triton X 100 and CTAB was remarkably enhanced by 6-folds and 3-folds, respectively which were remarkably higher in comparison to free immobilized lipase. The fabricated nanobiocatalysts were employed to synthesise green apple flavour ester, ethyl valerate via esterification reaction. Triton X 100 stabilized immobilized lipase was a better performer in yielding green apple flavour ester, demonstrating about 90% ester yield as compared to 78% yield obtained by CTAB stabilized immobilized lipase preparation. The obtained outcomes suggested that enzyme structure was stabilized by the GA treatment if executed in the absence or in the presence of detergent, and that, in the company of detergent, a conformation of the lipase with the exposed active center to the medium provided an aggrandized catalytic performance.

Structural-Dependent N,O-Donor Imine-Appended Cu(II)/Zn(II) Complexes: Synthesis, Spectral, and in Vitro Pharmacological Assessment.

Four mononuclear bioefficient imine-based coordination complexes, [(L 1 ) 2 Cu], [(L 1 ) 2 Zn], [(L 2 )Cu(H 2 O)], and [(L 2 )Zn(H 2 O)], were synthesized using ligands [L 1 = 2-(((3-hydroxynaphthalen-2-yl)methylene)amino)-2-methylpropane-1,3-diol and L 2 = 4-(1-((1,3-dihydroxy-2-methylpropan-2-yl)imino)ethyl)benzene-1,3-diol]. The formation of the complexes was ascertained by elemental analysis, Fourier transform infrared, 1H NMR, 13C NMR, electrospray ionization-mass spectroscopy, electron paramagnetic resonance, and thermogravimetric analysis. The comparative binding propensity profiles of the above-synthesized complexes with the DNA/human serum albumin (HSA) were investigated via UV absorption, fluorescence, and Förster resonance energy-transfer studies. On the basis of extended conjugation and planarity, L 1 complexes exhibited superior bioactivity with greater calculated DNA binding constant values, (K b) 2.9444 × 103 [(L 1 ) 2 Cu] and 2.2693 × 103 [(L 1 ) 2 Zn], as compared to L 2 complexes, 1.793 × 103 [(L 2 )Cu(H 2 O)] and 9.801 × 102 [(L 2 )Zn(H 2 O)]. The competitive displacement assay of complexes was performed by means of fluorogenic dyes (EtBr and Hoechst), which corroborates the occurrence of minor groove binding because of the enhanced displacement activity with Hoechst 33258. The minor groove binding of the [(L 1 ) 2 Cu] complex is further confirmed by the molecular docking study. Moreover, the HSA study demonstrated effective static quenching of complexes with substantial K sv values. The [(L 1 ) 2 Cu] complex was found to have pronounced cleavage efficiency as evaluated from sodium dodecyl sulfate polyacrylamide gel electrophoresis electrophoresis. Furthermore, in vitro antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl and superoxide radicals further proclaimed the remarkable bioefficiency of compounds, which make them promising as active chemotherapeutic agents.

Immobilization of lipase onto novel constructed polydopamine grafted multiwalled carbon nanotube impregnated with magnetic cobalt and its application in synthesis of fruit flavours.

Surface modification of multiwalled carbon nanotubes (MWCNTs) could enhance the features of the nanomaterial as carrier for enzyme immobilization. In this strategy, magnetic MWCNTs were fabricated by incorporating them with cobalt and functionalization was carried out by aminated polydopamine. The surface modified MWCNTs were then used as a carrier for the immobilization of Candida rugosa lipase (CRL) via covalent binding using glutaraldehyde. The immobilized CRL maintained high activity, which was 3-folds of free CRL. The immobilized CRL exhibited excellent thermal resistance as validated by TGA and DTA technique and was found to be active in a broad range of pH and temperatures in comparison to free CRL. Systematic characterization via FT-IR spectroscopy, CD spectroscopy, SEM, TEM and confocal laser scanning microscopy confirmed the presence of CRL on the modified MWCNTs. Immobilized CRL presented an exquisite recycling performance as after ten consecutive reuses it retained around 84% of its initial hydrolytic activity and further showed high yield enzymatic synthesis of ethyl butyrate and isoamyl acetate having characteristic pineapple and banana flavour demonstrating 78% and 75% ester yield, respectively. The present work provides a novel perspective for lipase catalyzed biotechnological applications by adding a magnetic gain to intrinsic features of MWCNTs.

A robust nanobiocatalyst based on high performance lipase immobilized to novel synthesised poly(o-toluidine) functionalized magnetic nanocomposite: Sterling stability and application.

Herein, as a promising support, a magnetic enzyme nanoformulation have been designed and fabricated by a poly-o-toluidine modification approach. Owing to the magnetic nature and the existence of amine functionalized groups, the as-synthesised poly(o-toluidine) functionalized magnetic nanocomposite (Fe3O4@POT) was employed as potential support for Candida rugosa lipase (CRL) immobilization to explore its application in fruit flavour esters synthesis. The morphology and structure of the Fe3O4@POT NC were examined through various analytical tools. Hydrolytic activity assays disclose that immobilized lipase demonstrated an activity yield of 120%. It is worth mentioning that CRL#Fe3O4@POT showed superior resistance to extremes of temperature and pH and different organic solvents in contrast to free CRL. The magnetic behaviour of the as-synthesised NC was affirmed by alternating gradient magnetometer analysis. It was found to own facile immobilization process, enhanced catalytic performance for the immobilized form which may be stretched to the immobilization of various vital industrial enzymes. Moreover, it retained improved recycling performance. After 10 cycles of repetitive uses, it still possessed around 90% of its initial activity for the hydrolytic reaction, since the enzyme-magnetic nanoconjugate was effortlessly obtained using a magnet from the reaction system. The formulated nanobiocatalyst was selected for the esterification reaction to synthesize the fruit flavour esters, ethyl acetoacetate and ethyl valerate. The immobilized lipase successfully synthesised flavour compounds in aqueous and n-hexane media having significant higher ester yields compared to free enzyme. The present work successfully combines an industrially prominent biocatalyst, CRL, and a novel magnetic nanocarrier, Fe3O4@POT, into an immobilized nanoformulation with upgraded catalytic properties which has excellent potential for practical industrial implications.

Synthesis, Characterization, and Applications of Nanographene-Armored Enzymes.

The unexpected discovery of graphene and especially the follow-up explosion of interest in its properties and applications marked the beginning of a new carbon era. Graphene-based nanostructured materials are highly useful because they show great promise in the field of biotechnology and biomedicine. Owing to their unique structural features, exceptional chemical, electrical, and mechanical properties, and their ability to affect the microenvironment of biomolecules, graphene-armored nanomaterials are suitable for use in various applications, such as immobilization of enzymes, field-effect transistors, photovoltaic devices, and biosensors, which in turn is extremely vital to the development of biomedical instruments, clinical diagnosis, and disease treatment. In this chapter, we present our recently reported work to armor hydrolytic enzymes on graphene-based nanomaterial in order to develop novel scaffolds to build robust nanobiocatalysts. Synthesized graphene-Fe3O4 and polyaniline-coated silver graphene nanocomposite have been used to immobilize ß-galactosidase and lipase, using noncovalent and covalent strategies, respectively. Herein, the methodologies of both techniques have been discussed in detail. Owing to the large surface area offered by the honeycomb like structure of graphene, very high amount of enzyme has been loaded on small amounts of the nanocomposite. The stability and reusability of the fabricated nanobiocatalysts have been compared with their free forms. Nanographene-armored enzymes demonstrated high catalytic stability and easy recovery from the reaction medium and can be applied in various biotechnological applications. Lastly, future prospects and possible challenges in this rapidly developing area have also been discussed.

Exquisite stability and catalytic performance of immobilized lipase on novel fabricated nanocellulose fused polypyrrole/graphene oxide nanocomposite: Characterization and application.

This work was performed to describe the facile procedure of a novel nanobiocatalyst, nano cellulose fused polypyrrole/graphene oxide nanocomposite for the efficacious immobilization of lipase, a versatile hydrolytic enzyme having potential applications in industries. The fabricated nanocomposite was characterized using Fourier transform infrared spectroscopy, differential thermal analysis, thermogravimetric analysis, X-ray diffraction, scanning electron microscopy, atomic force microscopy, transmission electron microscopy, and Candida rugosa lipase was immobilized onto nanocomposite through physical adsorption. The catalytic efficiency and operational stabilities of immobilized lipase were improved significantly compared to the free lipase. The reusability profile outcomes showed that the immobilized lipase formulation was an outstanding nanobiocatalyst as it retained 85% of its original catalytic activity after 10 cycles of application. The nanobiocatalyst was employed for the synthesis of the fruit flavour compound, ethyl acetoacetate. The immobilized lipase successfully synthesised flavour compound in solvent free media and n-hexane having 27.5% and 75.5% ester yields respectively. Moreover, these outcomes demonstrating graphene oxide modified carrier induced stabilization, amended solvent tolerance and operational stability of immobilized enzyme, will have quintessential influence on practical scale up of biotechnological industries.

Unraveling Comparative Anti-Amyloidogenic Behavior of Pyrazinamide and D-Cycloserine: A Mechanistic Biophysical Insight.

Amyloid fibril formation by proteins leads to variety of degenerative disorders called amyloidosis. While these disorders are topic of extensive research, effective treatments are still unavailable. Thus in present study, two anti-tuberculosis drugs, i.e., pyrazinamide (PYZ) and D-cycloserine (DCS), also known for treatment for Alzheimer's dementia, were checked for the anti-aggregation and anti-amyloidogenic ability on Aß-42 peptide and hen egg white lysozyme. Results demonstrated that both drugs inhibit the heat induced aggregation; however, PYZ was more potent and decelerated the nucleation phase as observed from various spectroscopic and microscopic techniques. Furthermore, pre-formed amyloid fibrils incubated with these drugs also increased the PC12/SH-SY5Y cell viability as compare to the amyloid fibrils alone; however, the increase was more pronounced for PYZ as confirmed by MTT assay. Additionally, molecular docking study suggested that the greater inhibitory potential of PYZ as compare to DCS may be due to strong binding affinity and more occupancy of hydrophobic patches of HEWL, which is known to form the core of the protein fibrils.