Characterization of a conjugate between poly(N-vinyl caprolactam) and a triazine-based covalent organic framework as a potential biomaterial.

Thermoresponsive polymers (TRPs) have been explored over decades for biomedical applications, and poly(N-vinylcaprolactam) (PVCL) TRP is extensively investigated due to its low toxicity and lower critical solution temperature (LCST), close to physiological temperatures. Besides this, the utilization of covalent organic frameworks (COFs), which belong to a class of porous polymers, in bio-based applications is of great interest due to their remarkable properties. Thus, the integration of PVCL and covalent organic frameworks (COFs) as conjugate materials can lead to advanced bio-based applications; however, the need is to understand the influence of the COF on the PVCL conformation. Herein, a triazine-based COF, CC-TAPT-COF, has been synthesized and completely characterized. Later, the effect of CC-TAPT-COF on the PVCL polymer conformation was studied using various techniques. In fluorescence spectroscopy, a fluorescence quenching for PVCL in the presence of CC-TAPT-COF was observed, which indicated conformational changes. Later, results from thermal fluorescence studies and dynamic light scattering as a function of temperature showed a slight decrease in LCST value for PVCL after the addition of CC-TAPT-COF concentrations. These results showed a slight effect of CC-TAPT-COF on the PVCL conformation. Likewise, a slight decrease in the transmittance value for specific bands in infrared spectra showed a slight effect of CC-TAPT-COF on the PVCL conformation. Further, results from electron microscopy and atomic force microscopy revealed a conjugate formation between PVCL and CC-TAPT-COF due to the presence of binding interactions between them. Overall, the results from several studies showed a slight effect of CC-TAPT-COF on the PVCL during conjugate formation between PVCL and CC-TAPT-COF. This study will be beneficial for the development of COF-thermoresponsive polymer conjugates with a mixture of their unique features as advanced biomaterials.

Sustainable combination of ionic liquid and deep eutectic solvent for protecting and preserving of the protein structure: The synergistic interaction of enzymes and eco-friendly hybrid ionic fluids.

Hybrid ionic fluids (HIFs) are one of the emerging and fascinating sustainable solvent media, a novel environment-friendly solvent for biomolecules. The HIFs have been synthesized by combining a deep eutectic solvent (DES), an ionic liquid (IL) having a common ion. The stability and activity of hen's egg white lysozyme (Lyz) in the presence of a recently designed new class of biocompatible solvents, HIFs have been explored by UV-visible, steady-state fluorescence, circular dichroism (CD), Fourier transform infrared spectroscopy (FT-IR) along with dynamic light scattering (DLS) measurements. This work emphasizes the effect of DES synthesized by using 1:2 choline chloride and glycerol [Glyn], ILs (1-butly-3-methylimidazolium chloride [BMIM]Cl and choline acetate [Chn][Ac]) and their corresponding HIFs on the structure and functionality of Lyz. Moving forward, we also studied the secondary structure, thermal stability and enzymatic activity and thermodynamic profile of Lyz at pH = 7 in the presence of varying concentrations (0.1 to 0.5) M of [BMIM]Cl, [Chn][Ac] ILs, [Glyn] DES and [Glyn][BMIM]Cl (hybrid ionic fluid1) as well as [Glyn][Chn][Ac] (hybrid ionic fluid2). Spectroscopic results elucidate that ILs affect the activity and structural stability of Lyz, whereas the stability and activity are increased by DES and are maintained by HIFs at all the studied concentrations. Overall, the experimental results studied elucidate expressly that the properties of Lyz are maintained in the presence of hybrid ionic fluid1 while these properties are intensified in hybrid ionic fluid2. This work has elucidated expressly biocompatible green solvents in protein stability and functionality due to the alluring properties of DES, which can counteract the negative effect of ILs in HIFs.

An eminent approach towards next generation solvents for sustainable packaging and stability of enzymes: a comprehensive study of ionic liquid and deep eutectic solvent mixtures.

Hybrid ionic fluids (HIFs) are newly emerging and fascinating sustainable solvent media, which are attracting a great deal of scientific interest in protecting the native structure of proteins. For a few decades, there has been a demand to consider ionic liquids (ILs) and deep eutectic solvents (DESs) as biocompatible solvent media for enzymes; however, in some cases, these solvent media also show limitations. Therefore, this work focuses on synthesising novel HIFs to intensify the properties of existing ILs and DESs by mixing them. Herein, HIFs have been synthesised by the amalgamation of a deep eutectic solvent (DES) and an ionic liquid (IL) with a common cation or anion. Later on, the stability and activity of hen's egg white lysozyme (Lyz) in the presence of biocompatible solvent media and HIFs were studied by various techniques such as UV-vis, steady-state fluorescence, circular dichroism (CD), Fourier transform infrared spectroscopy (FT-IR) and dynamic light scattering (DLS) measurements. This work emphasises the effect of a DES (synthesised using 1 : 2 choline chloride and malonic acid) [Maline], ILs (1-butyl-3-methylimidazolium chloride [BMIM]Cl or choline acetate [Chn][Ac]) and their corresponding HIFs on the structure and functionality of Lyz. Moreover, we also studied the secondary structure, thermal stability, enzymatic activity and thermodynamic profile of Lyz at pH = 7 in the presence of varying concentrations (0.1 to 0.5 M) of [BMIM]Cl and [Chn][Ac] ILs, Maline as a DES, and Maline [BMIM]Cl (HIF1) and Maline [Chn][Ac] (HIF2). Spectroscopic results elucidate that ILs affect the activity and structural stability of Lyz. In contrast, the stability and activity are inhibited by DES and are enhanced by HIFs at all the studied concentrations. Overall, the experimental results studied explicitly elucidate that the structure and stability of Lyz are maintained in the presence of HIF1 while these properties are intensified in HIF2. This study shows various applications in biocompatible green solvents, particularly in the stability and functionality of proteins, due to their unique combination where the properties counteract the negative effect of either DESs or ILs in HIFs.

Unraveling the Role of Deep Eutectic Solvents with Varying Hydrogen-Bond Acceptors on the Thermoresponsive Polymer Poly(N-isopropylacrylamide).

Deep eutectic solvents (DESs) have emerged as promising tools for crafting polymeric materials across diverse domains. This study delves into the impact of a series of DESs on the phase behavior of poly(N-isopropylacrylamide) (PNIPAM) in aqueous environments, presenting compelling insights into their performance. Specifically, we explore the conformational phase behavior of PNIPAM in the presence of four distinct lactic acid (LA)-based DESs: LA-betaine (LA-BET), LA-proline (LA-PRO), LA-choline chloride (LA-CC), and LA-urea (LA-U). By maintaining a consistent hydrogen-bond donor (HBD) while varying the hydrogen-bond acceptor (HBA), we unravel how different DES compositions modulate the phase transition behavior of PNIPAM. Our findings underscore the profound influence of DESs comprising LA as the HBD and diverse HBAs-BET, PRO, CC, and U on the thermoresponsive behavior of PNIPAM. Employing spectroscopic techniques such as ultraviolet-visible (UV-vis) spectroscopy, steady-state fluorescence, Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), ζ-potential, and transmission electron microscopy (TEM), we elucidate the preferential interactions between the HBA groups within DESs and the hydration layer of PNIPAM. Notably, temperature-dependent DLS analyses reveal a discernible decrease in the lower critical solution temperature (LCST) of PNIPAM with increasing DES concentration, ultimately disrupting the hydrogen-bond interactions and resulting in early hydrophobic collapse of the polymer, which can be clearly seen in the TEM micrographs. Furthermore, the formation of polymer composites within the mixed system leads to notable alterations in the physiochemical properties of PNIPAM, as evidenced by shifts in its LCST value in the presence of DESs. This perturbation disrupts hydrogen-bond interactions, inducing hydrophobic collapse of the polymers, a phenomenon vividly captured in TEM micrographs. In essence, our study sheds new light on the pivotal role of varying HBA groups within DESs in modulating the conformational transitions of PNIPAM. These insights not only enrich our fundamental understanding but also hold immense promise for the development of smart polymeric systems with multifaceted applications spanning bioimaging, biomedical science, polymer science, and beyond.

Green Synthesis of Silver Nanoparticles Using Drymaria cordata and Their Biocompatibility with Hemoglobin: A Therapeutic Potential Approach.

In this study, we present the synthesis and characterization of AgNPs using Drymaria cordata along with an assessment of their antioxidant, antibacterial, and antidiabetic activities. Antibacterial activities using four bacterial strains, free radical scavenging assays (DPPH and ABTS), and carbohydrate hydrolyzing enzyme inhibition assays were done to examine the therapeutic efficacy of AgNPs. Additionally, herein, we also evaluated the biocompatibility of the AgNPs using hemoglobin (Hb) as a model protein. A comprehensive analysis of Hb and AgNP interactions was carried out by using various spectroscopic, imaging, and size determination studies. Spectroscopic results showed that the secondary structure of Hb was not altered after its interaction with AgNPs. Furthermore, the thermal stability was also well maintained at different concentrations of nanoparticles. This study demonstrated a low-cost, quick, and eco-friendly method for developing AgNPs using D. cordata, and the biocompatible nature of AgNPs was also established. D. cordata-mediated AgNPs have potential applications against bacteria and diabetes and may be utilized for targeted drug delivery.

Impact of Choline Hydroxide-Supported Magnetic Nanoparticles on Peroxidase Activity and Conformational Stability of Cytochrome c.

Nanotechnology has advanced significantly; however, little is known about the potential implications on human health-related issues, particularly blood carrying enzymes. Ionic liquids are also well-recognized for maintaining the structure and activity of enzymes. In this regard, we delineate a facile synthetic approach of preparation of Fe3O4 nanoparticles (NPs) as well as choline hydroxide [CH][OH] ionic liquid (IL)-supported Fe3O4 NPs (Fe3O4-CHOH). This approach of combining magnetic nanoparticles (MNPs) with IL results in distinctive properties, which may offer enormous utility in the field of biomedical research due to the effortless separation of MNPs by an external magnetic field. Detailed characterization of MNPs including Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Raman spectroscopy, transmission electron microscopy (TEM), and scanning electron microscopy (SEM) was carried out. The biomolecular interactions of Fe3O4 and Fe3O4-CHOH NPs with cytochrome c (Cyt c) were studied in detail using various spectroscopic and microscopic techniques. From spectroscopic studies, it can be concluded that the secondary structure of Cyt c is more stable in the presence of Fe3O4-CHOH NPs than Fe3O4 NPs. The binding constant of Cyt c in the presence of MNPs was also calculated using the Benesi-Hildebrand equation. Furthermore, dynamic light scattering (DLS), ζ-potential, and microscopic studies were performed to study the interaction of Cyt c with MNPs. These studies provided evidence favoring the formation of bionanoconjugates of Cyt c with MNPs. Moreover, the enzymatic activity of Cyt c increases in the presence of both MNPs. The peroxidase activity of Cyt c in MNPs explicitly elucidates that the enzyme is preserved for a long time in the presence of Fe3O4-CHOH NPs. Later on, TEM and field emission scanning electron microscopy (FESEM) were also performed to gather more information regarding the morphology of Cyt c in the presence of MNPs.

Deep eutectic solvents induced changes in the phase transition behavior of smart polymers: a sustainable future approach.

Deep eutectic solvents (DESs) are considered "green" and "sustainable" alternatives to conventional organic solvents and ionic liquids (ILs) due to their characteristic properties and relatively low costs. DESs are considered IL analogs and have attracted consideration as benign media formulations for the synthesis of novel polymers because they satisfy the principle of sustainability. Over the past few years, the use of DESs has resulted in novel pathways for the synthesis of novel materials, biomaterials, functional materials, and ionic soft materials. Furthermore, DESs have been widely applied in the science, industrial, engineering, and technological fields. On the other hand, stimulus-responsive (smart) polymers have been widely utilized in intelligent devices owing to their virtues of good processibility, stimuli and environmental sensitivity, responsivity, and so on. With the introduction of a DES into the smart polymeric matrices, their potential characteristics, biocompatibility, and flexibility endow the corresponding DES-based polymeric materials with intriguing properties, which in turn will broaden their applications in various domains of polymer science and material chemistry. Substantial research has been done in the fabrication of DES-based polymeric materials. Numerous studies have extensively investigated the effects of DESs on biomolecules such as proteins/enzymes and nucleic acids, whereas few have addressed the impact of DESs on the aggregation and phase transition behaviors of smart polymers. This review focuses on mechanistic insights, aggregation behavior, and interactions between smart polymers and DESs. Opportunities and future research perspectives in this blossoming arena are also discussed. It is hoped that this review will pave futuristic pathways for the design and development of advanced DES-based polymeric materials and biomaterials for various applications.

Microplastics and biobased polymers to combat plastics waste.

Microplastics (MPs) have become the major global concern due to their adverse effects on the environment, human health, and hygiene. These complex molecules have numerous toxic impacts on human well-being. This review focuses on the methods for chemically quantifying and identifying MPs in real-time samples, as well as the detrimental effects resulting from exposure to them. Biopolymers offer promising solutions for reducing the environmental impact caused by persistent plastic pollution. The review also examines the significant progress achieved in the preparation and modification of various biobased polymers, including polylactic acid (PLA), poly(ε-caprolactone) (PCL), lignin-based polymers, poly-3-hydroxybutyrate (PHB), and poly(hydroxyalkanoates) (PHA), which hold promise for addressing the challenges associated with unplanned plastic waste disposal.

Assessing the compatibility of choline-based deep eutectic solvents for the structural stability and activity of cellulase: Enzyme sustain at high temperature.

As a new generation of 'green solvents' deep eutectic solvents (DESs) represents a promising alternative to the conventional solvents. Their environmental-benign nature and designer properties promote their utility in biocatalysis. Enzymes are marginally stable when exposed to physical/chemical disturbances. One such enzyme is cellulase which is a propitious catalyst for the depolymerization of cellulose under mild conditions. Therefore, their stability is a prerequisite condition to match demands of biorefineries. To address this issue of low stability, activity and thermal denaturation of cellulase, there is a need to find a sustainable and suitable co-solvent that is biocompatible with enzymes ultimately to facilitate their application in bio-industries. In this regard, we synthesized three choline-based DESs, choline chloride (ChCl)-glycerol, ChCl-ethylene glycol and ChCl-lactic acid and employed them to analyze their suitability for cellulase. The present study systematically evaluates the influence of the mentioned DESs on stability, activity and thermal stability of cellulase with the help of various spectroscopic techniques. The spectroscopic analysis revealed that the structural stability and activity of the enzyme were improved in presence of ChCl-glycerol and ChCl-ethylene glycol. The thermal stability was also very well maintained in both the DESs. Interestingly, the relative activity of cellulase was >80 % even after incubation at 50 °C after 48 h for both the DESs. This activity preservation behaviour was more pronounced for ChCl-ethylene glycol than ChCl-glycerol. Moreover, temperature variations studies also reveal promising results by maintain conformational intactness. On the other side, ChCl-lactic acid showed a deleterious effect on the enzyme both structurally as well as thermally. The dynamic light scattering (DLS) analysis provides more specific information about the negative influence of ChCl-lactic acid towards cellulase native structure. This DES induces unavoidable alterations in the enzyme structure which leads to the unfolding of enzyme, ultimately, destabilizing it. Overall, our results present a physical insight into how the enzyme stability and activity depend on the nature of DES. Also, the findings will help to facilitate the development and application of DESs as biocatalytic process.

The attenuating ability of deep eutectic solvents towards the carboxylated multiwalled carbon nanotubes induced denatured ß-lactoglobulin structure.

The stabilization of proteins has been a major challenge for their practical utilization in industrial applications. Proteins can easily lose their native conformation in the presence of denaturants, which unfolds the protein structure. Since the introduction of deep eutectic solvents (DESs), there are numerous studies in which DESs act as promising co-solvents that are biocompatible with biomolecules. DESs have emerged as sustainable biocatalytic media and an alternative to conventional organic solvents and ionic liquids (ILs). However, the superiority of DESs over the deleterious influence of denaturants on proteins is often neglected. To address this, we present the counteracting ability of biocompatible DESs, namely, choline chloride-glycerol (DES-1) and choline chloride-urea (DES-2), against the structural changes induced in ß-lactoglobulin (Blg) by carboxylated multiwalled carbon nanotubes (CA-MWCNTs). The work is substantiated with various spectroscopic and thermal studies. The spectroscopic results revealed that the fluorescence emission intensity enhances for the protein in DESs. Contrary to this, the emission intensity extremely quenches in the presence of CA-MWCNTs. However, in the mixture of DESs and CA-MWCNTs, there was a slight increase in the fluorescence intensity. Circular dichroism spectral studies reflect the reappearance of the native band that was lost in the presence of CA-MWCNTs, which is a good indicator of the counteraction ability of DESs. Further, thermal fluorescence studies showed that the protein exhibited extremely great thermal stability in both DESs as well as in the mixture of DES-CA-MWCNTs compared to the protein in buffer. This study is also supported by dynamic light scattering and zeta potential measurements; the results reveal that DESs were successfully able to maintain the protein structure. The addition of CA-MWCNTs results in complex formation with the protein, which is indicated by the increased hydrodynamic size of the protein. The presence of DESs in the mixture of CA-MWCNTs and DESs was quite successful in eliminating the negative impact of CA-MWCNTs on protein structural alteration. DES-1 proved to be superior to DES-2 over counteraction against CA-MWCNTs and maintained the native conformation of the protein. Overall, both DESs act as recoiling media for both native and unfolded (denatured by CA-MWCNTs) Blg structures. Both the DESs can be described as potential co-solvents for Blg with increased structural and thermal stability of the protein. To the best of our knowledge, this study for the first time has demonstrated the role of choline-based DESs in the mixture with CA-MWCNTs in the structural transition of Blg. The DESs in the mixture successfully enhance the stability of the protein by reducing the perturbation caused by CA-MWCNTs and then amplifying the advantages of the DESs present in the mixture. Overall, these results might find implications for understanding the role of DES-CA-MWCNT mixtures in protein folding/unfolding and pave a new direction for the development of eco-friendly protein-protective solvents.

Impact of ZIF-8, ArgHCl, and Ionic Liquid-Based Formulations on the Conformational and Colloidal Stability of Antibodies.

Various formulations consisting of biomaterials zirconium imidazolate framework-8 (ZIF-8), choline acetate ([Ch][Ac]), and arginine hydrochloride (argHCl) are optimized to study the stability of antibody, Immunoglobulin G (IgG). We have performed several instrumentations including UV-visible spectroscopy, dynamic light scattering (DLS), circular dichroism (far UV CD), and atomic force microscopy (AFM) in the presence of all the formulations to investigate the conformational and colloidal stability of the antibodies. Alongside, the packing efficiency of all the formulations was also explored by storing IgG at 4 °C for 3 months. We have tried to investigate the interactions between biomaterials and antibodies with the motive of designing aggregation-resistant formulations. The overall stability of IgG was improved in the presence of [Ch][Ac]; however, ZIF-8 and argHCl cause relatively more aggregation, although the structure was retained in all the formulations. The key aspect of this study is that the presence of [Ch][Ac] increases ZIF-8@IgG's thermal stability and resistance to IgG-argHCl aggregation. All over, for the first time, with different experimental approaches, the impact of each biomaterial individually and in combination is explored to study their effect on the stability of antibodies. Thus, better efficient formulations can be designed for the storage/packaging of IgG-based drugs which ultimately will have more applicability in pharmaceuticals.

Ionic Liquids Mediated Micellization of Pluronic Copolymers: Aggregation Behavior of Amphiphilic Triblock Copolymers.

Understanding the micellization of amphiphilic triblock copolymers, especially Pluronics can play a persuasive role in engineering "smart" formulations for drug delivery applications. Their underlying self-assembly in the presence of designer solvents such as ionic liquids (ILs) provides combinatorial benefits of unique munificent properties of ILs and copolymers. The complex molecular interactions in the Pluronic copolymers/ILs mixed system influence the aggregation mechanism of copolymers depending on various aspects with no standardized factors to govern the structure-property relationship, which led to the practical applications. Here, we summarized recent progress in understanding the micellization process of IL-Pluronic mixed systems. Special emphasis was given to pure Pluronic systems (i.e., PEO-PPO-PEO) without any structural modifications, such as copolymerization with other functional groups, and ILs having cholinium and imidazolium groups. We expect that the correlation between existing/developing experimental and theoretical studies will provide the necessary basis and impetus for successful utilization in drug delivery applications.

Smart Anisotropic Colloidal Composites: A Suitable Platform for Modifying the Phase Transition of Diblock Copolymers by Gold Nanoparticles.

Surface modification of metallic nanoparticles (NPs) by stimuli-responsive polymers is a benign method to prepare smart colloidal composites which tune the characteristic properties of individual systems. The temperature-dependent transition of diblock copolymer poly(N-isopropylacrylamide)-block-poly(N-vinylcaprolactam) (PNIPMA-b-PVCL) synthesized using reversible addition-fragmentation chain transfer polymerization was studied by incorporating anisotropic gold NPs (AGPs) such as spheres (AuNSs), rods (AuNRs), cubes (AuNCs), and rhombic dodecahedrals (AuRDs). Shape-dependent physiochemical properties of nanostructures alter the lower critical solution temperature (LCST) of the chemical inhomogeneous diblock copolymer. Heterogeneous nucleation of AuNPs was facilitated by seed-mediated synthesis for incorporating uniformity. In the mixed system, the presence of PNIPAM-b-PVCL modifies the surface of AGPs through physisorption which is supported by transmission electron microscopy and field emission scanning electron microscopy showing the NPs embedding in the polymeric matrix. Furthermore, steady state fluorescence spectroscopy and Fourier transform infrared spectroscopy were performed to examine the phase transition behavior of PNIPAM-b-PVCL in AGPs. The formation of a smart polymer nanocomposite alters the physiochemical properties of the diblock copolymer as demonstrated from the variation of LCST in the dynamic light scattering measurement. Henceforth, functionalizing the surfaces of AGPs with a thermoresponsive diblock copolymer provides combinatorial benefits in the properties of smart polymeric colloidal systems with potential applications in bioimaging and drug delivery.

Fortification of thermal and structural stability of hemoglobin using choline chloride-based deep eutectic solvents.

Of late, DESs have occupied the centre stage due to their eco-friendly and resource-efficient nature and their low toxicity. In this work, we have investigated the structural and thermal stability of hemoglobin (Hb) in two choline chloride ([Ch]Cl)-based DESs namely urea [Ch]Cl-urea (DES1) and [Ch]Cl-glycerol (Gly); (DES 2). Different biophysical techniques reveal that the presence of DESs facilitates the stability of Hb in a concentration-dependent manner and the extent of stability is more pronounced in [Ch]Cl-Gly as compared to [Ch]Cl-urea. Additionally, for a better understanding of the role of DESs in modulating the thermal and structural stability of Hb, studies have been performed on Hb in the presence of individual constituents of DESs, i.e., [Ch]Cl, urea, and Gly. Altogether, it was observed that the effect on the stability of Hb was by the presence of the DESs rather than their individual constituents. For instance, urea itself is a destabilizing co-solvent for biomolecules. However, the harmful effects of urea were surpassed when a DES is formed in the presence of [Ch]Cl. Therefore, overall, it can be concluded that both DESs can be described as potential non-harmful, green, and promising solvents for enhancing the structural and thermal stability of Hb.

Effect of Imidazolium Nitrate Ionic Liquids on Conformational Changes of Poly(N-vinylcaprolactam).

Detailed information about molecular interactions and conformational changes of polymeric components in the presence of ionic liquids (ILs) is essential for designing novel polymeric ionic liquid-based biomaterials. In biomaterials science and technology, thermoresponsive polymers (TRPs) are widely viewed as potential candidates for the fabrication of biorelated medical devices. Here, we synthesized thermoresponsive poly(N-vinyl-caprolactam) (PVCL) polymer and investigated the effects of imidazolium-based ILs (1-ethyl-3-methyl imidazolium nitrate and 1-butyl-3-methylimidazolium nitrate) with common anion and different cations on the phase transition behavior of PVCL aqueous solution. The impact of ILs on the phase transition behavior of PVCL was monitored by using UV-visible absorption spectra, steady-state fluorescence spectroscopy, thermal fluorescence spectroscopy, and temperature dependent dynamic light scattering. Results showed significant changes in the absorbance, molecular interactions, agglomeration, and coil to globule transition behaviors of PVCL in the presence of two ILs. PVCL aqueous solution showed significant conformational changes after the addition of ILs.

Unprecedented enhancement and preservation of the peroxidase activity of cytochrome-c packaged with ionic liquid-modified gold nanoparticles by offsetting temperature and time stresses.

Inspired by the biocompatibility of ionic liquids and their dexterousness for the preservation of enzyme structure and activity, herein, the interactions of Cyt-c with naked AuNPs and four IL-mediated AuNPs, which were formed by the fabrication of ILs with common cation 1-ethyl-3-methyl-imidazolium (EMIM) and different anions, to obtain AuNP-IL1 [(BF4)-1 anion], AuNP-IL2 [(CH3OSO3)-1 anion], AuNP-IL3 [(CH3CH2OSO3)-1 anion], and (AuNP-IL4) [Cl-1 anion], were studied. Through this work, the peroxidase activity observed in the presence of a lower concentration IL-AuNPs is exceptionally increased (16 fold). IL-AuNPs preferentially counteract the temperature gradient change and long-term solvent preservation effects while persistently maintaining the Cyt-c peroxidase activity without much depreciation. The hydrodynamic diameter (dH) of the Cyt-c-AuNP system was obtained, which supported the TEM results. Furthermore, to evaluate the effect of Cyt-c interaction with the AuNPs, a Zeta potential analysis was performed. Taken together, the binding of IL-AuNPs with Cyt-c, diameter size analysis, zeta potential, structural integrity evaluation using the DichroWeb software and morphology results suggest the interaction order of the IL-AuNPs to be in a sequence of AuNP-IL2 > AuNP-IL3 > AuNP- IL4 > AuNP-IL1 > Naked AuNPs. Moreover, results indicate that the IL anions play a dominating role in the modulation of interactions between IL-mediated AuNPs and Cyt-c. The study strongly supports the promising character of sulfur-containing IL-mediated AuNPs for Cyt-c immobilization simultaneously opening new avenues for the application of greener and biocompatible nanoparticles with drug delivery and therapeutic applications.

Current understanding and insights towards protein stabilization and activation in deep eutectic solvents as sustainable solvent media.

Deep eutectic solvents (DESs) have emerged as a new class of green, designer and biocompatible solvents, an alternative to conventional organic solvents and ionic liquids (ILs) which are comparatively toxic and non-biodegradable. DESs are eutectic mixtures that are formed when a hydrogen bond acceptor (HBA) is mixed with a hydrogen bond donor (HBD) at particular molar ratios by mechanical grinding or under mild heating conditions. Very recently, these solvents have been the center of attention for researchers in biotechnology, biomedicine and various scientific applications. These environmentally benign solvents have a close analogy with ILs; however, they offer certain unique merits over traditional ILs. DESs display remarkable properties such as easy preparation, tunable composition, biodegradability, recyclability, inherently low toxicity, sustainability and biocompatibility; these special features validate DESs as new potential solvents/co-solvents for biomolecules. Mechanistically, the biocompatibility and protein friendly nature of DESs depend on various factors, which include the composition of the DES, viscosity and hydration level. Therefore, it becomes an essential task to bring together all the studies related to protein behaviour in DESs to unlock their biomolecular proficiency. This review specifically highlights recent insights into the biomacromolecular functionality in DESs, including outlines of the solubilization and stabilization of proteins, long term protein packaging, different extraction methods and enzyme activation in the presence of DESs. A literature survey reveals that DESs act as green media in which the protein structure and activity are retained. In some cases, proteins refolded and enzymatic activity was enhanced several fold in the presence of DESs. Furthermore, we have reviewed the possible mechanistic behaviour behind protein stabilization, refolding and activation in DESs. Overall, the main objective of this review is to explicate the advantages of the introduction of DESs for biomolecules and to demonstrate the versatility of these eco-friendly solvents for future bio-based applications.

Role of protein-copolymer assembly in controlling micellization process of amphiphilic triblock copolymer.

HYPOTHESIS: Triblock copolymer poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol) (PEG-PPG-PEG) forms a well-known micellar assembly at a particular temperature. Apart from regular assembly within the copolymer, it is crucial to explore additional assembly behaviour via simple exposure of proteins which unveils biased interactions with blocks of copolymer. The current work focuses on the examination of Pluronic F108 i.e. PEG-PPG-PEG with two different proteins i.e. α-chymotrypsin (CT) and lysozyme (LSZ), aiming at probing the critical micellization temperature (CMT) and molecular level interactions. EXPERIMENTS: Potential role of protein-copolymer assembly formation at a particular concentration of protein in modulating CMT was shown by a systematic experimental approach combined with a series of physicochemical methods. The sophisticated multiple techniques include fluorescence spectroscopy, Fourier transform infrared (FTIR) spectroscopy, Raman spectroscopy, dynamic light scattering (DLS), zeta potential measurements, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Furthermore, molecular docking studies were also employed to correlate theoretical insights with experimental findings. FINDINGS: CT and LSZ decrease CMT in regular concentration-dependent manner except for particular concentration (1.5 mg/mL) of LSZ which shows anomalous behaviour in steady-state fluorescence spectroscopy, temperature dependent fluorescence spectroscopy, Raman spectroscopy and DLS measurements. SEM and TEM results clearly reveal protein-copolymer assembly formation. The assembled structure has different biophysical properties. Docking studies elucidate several bio macromolecular interactions which can be involved in assembly formation. Based on obtained results from biophysical techniques mechanism of CMT variation was deduced. Obtained results can be useful in biosensors and targeted drug delivery systems.

Understanding the close encounter of heme proteins with carboxylated multiwalled carbon nanotubes: a case study of contradictory stability trend for hemoglobin and myoglobin.

Carbon nanotubes (CNTs) are one of the unique and promising nanomaterials that possess plenty of applications, such as biosensors, advanced drug delivery systems and biotechnology. CNTs bind rapidly with proteins, which result in the formation of a protein coating layer known as a "protein corona" around the surface of the nanomaterial. This hinders their applications as a drug carrier and influences the properties of biological macromolecules. The present work focuses on studying the thermal stability and molecular level interactions of two heme proteins, hemoglobin (Hb) and myoglobin (Mb), in the presence of carboxylated functionalized multi-walled CNTs (CA-MWCNTs). Through the current study, the following steps have been taken to distinguish the biocompatibility of the hydrophilic surface CA-MWCNTs for heme proteins via a series of spectroscopic techniques and differential scanning calorimetry (DSC). UV-Visible and steady-state fluorescence spectroscopy were used to reveal changes in the aromatic amino acid residues of heme proteins upon the addition of CA-MWCNTs. Circular dichroism spectroscopy (CD) shows the alteration in the native structure of proteins in the presence of the nanomaterial. A tremendous increase in the size of the protein CA-MWCNTs system is observed in dynamic light scattering (DLS), which clearly manifests the protein corona formation. Unexpectedly, both proteins interact differently with CA-MWCNTs, which is observed in CD spectroscopy and DSC. In the presence of CA-MWCNTs, an increase in the transition temperature (Tm) was observed for Hb, while the Tm value decreases for Mb. Different interactions with proteins at the molecular scale may be the reason for this unexpected behavior. Henceforth, the present results can help in the design of the next-generation drug carrier nanomaterials with the idea of the heme protein corona formation prior to development.

The biocompatible validity of amino acid ionic liquid mediated gold nanoparticles for enhanced activity and structural stability of papain.

During the past few decades, gold nanoparticles (AuNPs) have attracted a lot of attention owing to their biomedical applications, like therapeutics and drug delivery; however, the detailed biomolecular interactions and structural alteration of naturally occurring biomolecules, such as enzymes, in AuNPs remain unknown. The effects of various additives on the thermal and structural properties, and activity of proteins/enzymes have been scavenged and communicated intensively in the literature; however, the synthesis of ionic liquid (IL) mediated AuNPs solely for the purpose of enzyme activity boosting and stability modulation has not yet been reported. In the current study, we explore the role of cholinium tryptophan [CHO][Trp] and tetraethyl tryptophan [TEA][Trp]IL-mediated gold nanoparticles (AuNPs) on the activity enhancement and structural stability of papain. Our results showed that [CHO][Trp] and [TEA][Trp]IL-mediated AuNPs efficiently increased the proteolytic activity of papain, which was increased from 100 to 206% for [CHO][Trp]IL-mediated AuNPs and enhanced from 100 to 136% in [TEA][Trp]IL-mediated AuNPs. Additionally, extended differential scanning calorimetry (DSC) results showed that these AAIL-mediated AuNPs maintained the thermal stability of papain only at lower concentration. Spectroscopic studies conclude that the tryptophan (Trp) group of papain is expanded more towards the polar environment in the presence of [CHO][Trp] as compared to [CHO][Trp]IL mediated AuNPs. The far CD spectral and deconvoluted results show that the α-helical and ß-turn contents of the secondary structure of papain are preserved to a large extent; however, disruption in the ß-sheet has been observed for both AAIL-mediated AuNPs. Dynamic light scattering (DLS), zeta potential and transmission electron microscopy (TEM) results illustrate the distinct interactive behavior of papain for both types of AAIL-mediated AuNPs. The immobilization of papain is higher on [CHO][Trp]AuNPs compared to [TEA][Trp]AuNPs and papain surrounds [CHO][Trp]AuNPs on all sides, which is lacking in [TEA][Trp]AuNPs.