Applications In Nanobiotechnology Research Articles

Triggered Reversible Disassembly of an Engineered Protein Nanocage*.

Protein nanocages play crucial roles in sub-cellular compartmentalization and spatial control in all domains of life and have been used as biomolecular tools for applications in biocatalysis, drug delivery, and bionanotechnology. The ability to control their assembly state under physiological conditions would further expand their practical utility. To gain such control, we introduced a peptide capable of triggering conformational change at a key structural position in the largest known encapsulin nanocompartment. We report the structure of the resulting engineered nanocage and demonstrate its ability to disassemble and reassemble on demand under physiological conditions. We demonstrate its capacity for in vivo encapsulation of proteins of choice while also demonstrating in vitro cargo loading capabilities. Our results represent a functionally robust addition to the nanocage toolbox and a novel approach for controlling protein nanocage disassembly and reassembly under mild conditions.

Triggered Reversible Disassembly of an Engineered Protein Nanocage**

AbstractProtein nanocages play crucial roles in sub‐cellular compartmentalization and spatial control in all domains of life and have been used as biomolecular tools for applications in biocatalysis, drug delivery, and bionanotechnology. The ability to control their assembly state under physiological conditions would further expand their practical utility. To gain such control, we introduced a peptide capable of triggering conformational change at a key structural position in the largest known encapsulin nanocompartment. We report the structure of the resulting engineered nanocage and demonstrate its ability to disassemble and reassemble on demand under physiological conditions. We demonstrate its capacity for in vivo encapsulation of proteins of choice while also demonstrating in vitro cargo loading capabilities. Our results represent a functionally robust addition to the nanocage toolbox and a novel approach for controlling protein nanocage disassembly and reassembly under mild conditions.

Antioxidant Activity of Bioactive Peptide Fractions from Germinated Soybeans Conjugated to Fe3O4 Nanoparticles by the Ugi Multicomponent Reaction.

The conjugation of biomolecules to magnetic nanoparticles has emerged as promising approach in biomedicine as the treatment of several diseases, such as cancer. In this study, conjugation of bioactive peptide fractions from germinated soybeans to magnetite nanoparticles was achieved. Different fractions of germinated soybean peptides (>10 kDa and 5–10 kDa) were for the first time conjugated to previously coated magnetite nanoparticles (with 3-aminopropyltriethoxysilane (APTES) and sodium citrate) by the Ugi four-component reaction. The crystallinity of the nanoparticles was corroborated by X-ray diffraction, while the particle size was determined by scanning transmission electron microscopy. The analyses were carried out using infrared and ultraviolet–visible spectroscopy, dynamic light scattering, and thermogravimetry, which confirmed the coating and functionalization of the magnetite nanoparticles and conjugation of different peptide fractions on their surfaces. The antioxidant activity of the conjugates was determined by the reducing power and hydroxyl radical scavenging activity. The nanoparticles synthesized represent promising materials, as they have found applications in bionanotechnology for enhanced treatment of diseases, such as cancer, due to a higher antioxidant capacity than that of fractions without conjugation. The highest antioxidant capacity was observed for a >10 kDa peptide fraction conjugated to the magnetite nanoparticles coated with APTES.

Crystalline S-Layer Protein Monolayers Induce Water Turbulences on the Nanometer Scale

Bacterial surface layers (S-layers) have been observed as the outermost cell envelope component in a wide range of bacteria and most archaea. They are one of the most common prokaryotic cell surface structures and cover the cells completely. It is assumed that S-layers provide selection advantages to prokaryotic cells in their natural habitats since they act as protective envelopes, as structures involved in cell adhesion and surface recognition, as molecular or ion traps, and as molecular sieves in the ultrafiltration range. In order to contribute to the question of the function of S-layers for the cell, we merged high-resolution cryo-EM and small-angle X-ray scattering datasets to build a coarse-grained functional model of the S-layer protein SbpA from Lysinibacillus sphaericus ATCC 4525. We applied the Navier–Stokes and the Poisson equations for a 2D section through the pore region in the self-assembled SbpA lattice. We calculated the flow field of water, the vorticity, the electrostatic potential, and the electric field of the coarse-grained model. From calculated local changes in the flow profile, evidence is provided that both the characteristic rigidity of the S-layer and the charge distribution determine its rheological properties. The strength of turbulence and pressure near the S-layer surface in the range of 10 to 50 nm thus support our hypothesis that the S-layer, due to its highly ordered repetitive crystalline structure, not only increases the exchange rate of metabolites but is also responsible for the remarkable antifouling properties of the cell surface. In this context, studies on the structure, assembly and function of S-layer proteins are promising for various applications in nanobiotechnology, biomimetics, biomedicine, and molecular nanotechnology.

Engineering and functional characterization of a proton-driven \u03b2-lactam antibiotic translocation module for bionanotechnological applications

Novel approaches in synthetic biology focus on the bottom-up modular assembly of natural, modified natural or artificial components into molecular systems with functionalities not found in nature. A possible application for such techniques is the bioremediation of natural water sources contaminated with small organic molecules (e.g., drugs and pesticides). A simple molecular system to actively accumulate and degrade pollutants could be a bionanoreactor composed of a liposome or polymersome scaffold combined with energizing- (e.g., light-driven proton pump), transporting- (e.g., proton-driven transporter) and degrading modules (e.g., enzyme). This work focuses on the engineering of a transport module specific for β-lactam antibiotics. We previously solved the crystal structure of a bacterial peptide transporter, which allowed us to improve the affinity for certain β-lactam antibiotics using structure-based mutagenesis combined with a bacterial uptake assay. We were able to identify specific mutations, which enhanced the affinity of the transporter for antibiotics containing certain structural features. Screening of potential compounds allowed for the identification of a β-lactam antibiotic ligand with relatively high affinity. Transport of antibiotics was evaluated using a solid-supported membrane electrophysiology assay. In summary, we have engineered a proton-driven β-lactam antibiotic translocation module, contributing to the growing toolset for bionanotechnological applications.

Control of foliar phytoparasitic nematodes through sustainable natural materials: Current progress and challenges.

Nematodes are hidden enemies that inhibit the entire ecosystem causing adverse effects on animals and plants, leading to economic losses. Management of foliar phytoparasitic nematodes is an excruciating task. Various approaches were used to control nematodes dispersal, i.e., traditional practices, resistant cultivars, plant extract, compost, biofumigants, induced resistance, nano-biotechnology applications, and chemical control. This study reviews the various strategies adopted in combating plant-parasitic nematodes while examining the benefits and challenges. The significant awareness of biological and environmental factors determines the effectiveness of nematode control, where the incorporation of alternative methods to reduce the nematodes population in plants with increasing crop yield. The researchers were interested in explaining the fundamental molecular mechanisms, providing an opportunity to deepen our understanding of the sustainable management of nematodes in croplands. Eco-friendly pesticides are effective as a sustainable nematodes management tool and safe for humans. The current review presents the eco-friendly methods in controlling nematodes to minimize yield losses, and benefit the agricultural production efficiency and the environment.

From Folding to Assembly: Functional Supramolecular Architectures of Peptides Comprised of Non‐Canonical Amino Acids

Front Cover: This review discusses the self-assembly of various noncoded amino acid-containing peptides and their applications in nanobiotechnology. The incorporation of non-coded amino acids in peptide sequences have been shown to form well defined secondary structures, similar to natural proteins, with the ability to self-assemble and generate a wide variety of nanostructures with excellent biocompatibility and biostability. This is reported by Rajkumar Misra, Safra Rudnick-Glick, and Lihi Adler-Abramovich in article number 2100090.

Recent nanobiotechnological advancements in lignocellulosic biomass valorization: A review

Increase in human population, rapid industrialization, excessive utilization of fossil fuel utilization and anthropogenic activities have caused serious threats to the environment in terms of greenhouse gas emissions (GHGs), global warming, air pollution, acid rain, etc. This destruction in sustainability can be averted by a paradigm shift in the fuel production from fossil resources to bioenergy. Amongst different forms of bioenergy, lignocellulosic biomass can be utilized as an attractive substrate for the production of several high-value products owing to its renewability, easy availability, and abundance. Additionally, utilization of these waste biomasses reduces the environmental hazards associated with its disposal. Impedance of lignin and crystalline nature of cellulose pose major bottlenecks in biomass based energy. Though, several physio-chemicals processes are recommended as mitigation route but none of them seems to be promising for large scale application. In recent years, a right fusion of biological treatment combined with nanotechnology for efficient pretreatment and subsequent hydrolysis of biomass by ubiquitous enzymes seems to be promising alternative. In addition, to overcome these difficulties, nanotechnology-based methods have been recently adopted in catalytic valorization of lignocellulosic biomass. The present review has critically discussed the application of nano-biotechnology in lignocellulosic biomass valorization in terms of pretreatment and hydrolysis. A detailed discussion on the application of various nanoparticles in these processes, enzyme immobilization and end-production utilization is presented in this review. Finally, the review emphasizes the major challenges of this process along with different routes and recommendations to address the issues.

Eco-Friendly Production of AuNPs and Their Impact on the Oil Oxidative Stability.

Vegetable oils have been used for different applications and, more recently, as an active host medium to obtain nanoparticles for employment in bionanotechnological applications. Nevertheless, oils are very susceptible to oxidation during production, storage, and transportation because of their chemical composition. Consequently, any modification in their production must be accompanied by an analysis of the oxidative stability. In this study, naked and biocompatible gold nanoparticles (AuNPs) were grown on sunflower oil during sputtering deposition using different deposition times. Size and morphology were determined by transmission electron microscopy (TEM) and their concentrations were found by inductively coupled plasma-optical emission spectroscopy (ICP-OES). Rancimat® method was employed to evaluate the AuNPs influence on the oxidative stability of the vegetable oil. Well-dispersed quasi-spherical NPs were produced with a mean diameter in the 2.9–3.7 nm range and they were concentration-dependent on the deposition time. A concentration of about 11 mg/L, 38 mg/L, and 225 mg/L of AuNPs was obtained for a deposition time of 5 min, 15 min, and 30 min, respectively. The results also revealed that AuNPs negatively affected the oxidative stability of the sunflower oil and exponentially reduced the induction period (IP) with the increase in AuNPs content. IP reductions of 63%, 77%, and 81% were determined for the AuNPs containing samples at 11 mg/L, 38 mg/L, and 225 mg/L. For the first time, it is reported that naked AuNPs promote the rapid degradation of vegetable oil and this points out the need for attention relative to the quality of vegetable oils used to host metal nanoparticles.

Physicochemical and antibacterial characterization of Aspergillus sp. filtrate-reduced graphene oxide

Graphene-based nanomaterials are rapidly emerging as antimicrobial agents against a range of pathogenic microorganisms. In the present study, we have demonstrated the synthesis of reduced graphene oxide (RGO) using Aspergillus sp. filtrate as a reducing agent. The obtained Aspergillus sp. filtrate-RGO (AF-RGO) was characterized by ultraviolet–visible absorption spectroscopy, dynamic light scattering, X-ray diffraction, Fourier-transform infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, and atomic force spectroscopy to obtain details about the size, composition, structure, and morphology. The characterization confirmed the reduction of graphene oxide, the restoration of sp2 conjugation, and the formation of few-layered RGO sheets. The surface charge and stability of the AF-RGO in aqueous media were examined by zeta potential measurements. The thermal stability of AF-RGO was increased significantly due to the restoration of the graphitic sp2 network. The antibacterial activity of AF-RGO was evaluated using the agar well diffusion method, broth microdilution method, and Live/Dead bacterial viability assay. AF-RGO exhibited good antibacterial activities against pathogenic Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of 15 and 30 mg/ml, respectively. Therefore, fungal filtrate-mediated synthesis of RGO is a promising antibacterial carbon nanomaterial for a range of biomedical and nano(bio)technological applications.

Silver nanoparticles stabilized by ramnolipids: Effect of pH

Rhamnolipids are glycolipid biosurfactants that have remarkable physicochemical characteristics, such as the capacity for self-assembly, which makes these biomolecules a promising option for application in nanobiotechnology. Rhamnolipids produced from a low-cost carbon source (glycerol) were used to stabilize silver nanoparticles. Silver nanoparticles (AgNPs) have been the subject of studies due to their physical chemical as well as biological properties, which corroborate their catalytic and antimicrobial activity. We compared nanoparticles obtained with three different pH values during synthesis (5, 7 and 9) in the presence of rhamnolipids. Dynamic light scattering showed that larger particles were formed at pH 5 (78–190 nm) compared to pH 7 (6.5–43 nm) and 9 (5.6–28.1 nm). Moreover, nanoparticle stability (analyzed based on the zeta potential) was enhanced with the increase in pH from 5 to 9 (−29.86 ± 1.04, −37.83 ± 0.90 and −40.33 ± 0.57 mV, respectively). Field emission gun scanning electron microscopy confirmed the round morphology of the silver nanoparticles. The LSPR spectra of AgNP for the pHs studied are conserved. In conclusion, different pH values in the presence of rhamnolipids used in the synthesis of silver nanoparticles directly affect nanoparticle size and stability.

Mechanistic Insights into Interactions at Urea-Hydroxyapatite Nanoparticle Interface.

Development of controlled release biomolecules by surface modification of hydroxyapatite nanoparticles has recently gained popularity in the areas of bionanotechnology and nanomedicine. However, optimization of these biomolecules for applications such as drug delivery, nutrient delivery requires a systematic understanding of binding mechanisms and interfacial kinetics at the molecular level between the nanomatrix and the active compound. In this research, urea is used as a model molecule to investigate its interactions with two morphologically different thin films of hydroxyapatite nanoparticles. These thin films were fabricated on quartz crystal piezoelectric sensors to selectively expose Ca2+ and PO43- sites of hydroxyapatite. Respective urea adsorption and desorption on both of these sites were monitored in situ and in real time in the phosphate buffer solution that mimics body fluids. The measured kinetic parameters, which corroborate structural predisposition for controlled release, show desorption rates that are one-tenth of the adsorption rates on both surfaces. Furthermore, the rate of desorption from the PO43- site is one-half the rate of desorption from the Ca2+ site. The Hill kinetic model was found to satisfactorily fit data, which explains cooperative binding between the hydroxyapatite nanoparticle thin film and urea. Fourier transform infrared spectra and X-ray photoemission spectra of the urea adsorbed on the above surfaces confirm the cooperative binding. It also elucidates the different binding mechanisms between urea and hydroxyapatite that contribute to the changes in the interfacial kinetics. These findings provide valuable information for structurally optimizing hydroxyapatite nanoparticle surfaces to control interfacial kinetics for applications in bionanotechnology and nanomedicine.

Current Status of Brain Tumor in the Kingdom of Saudi Arabia and Application of Nanobiotechnology for Its Treatment: A Comprehensive Review.

Objective: Brain tumors are the most challenging of all tumors and accounts for about 3% of all cancer allied deaths. The aim of the present review is to examine the brain tumor prevalence and treatment modalities available in the Kingdom of Saudi Arabia. It also provides a comprehensive analysis of the application of various nanotechnology-based products for brain cancer treatments along with their prospective future advancements. Methods: A literature review was performed to identify and summarize the current status of brain cancer in Saudi Arabia and the scope of nanobiotechnology in its treatment. Results: Depending upon the study population data analysis, gliomas, astrocytoma, meningioma, and metastatic cancer have a higher incidence rate in Saudi Arabia than in other countries, and are mostly treated in accordance with conventional treatment modalities for brain cancer. Due to the poor prognosis of cancer, it has an average survival rate of 2 years. Conventional therapy includes surgery, radiotherapy, chemotherapy, and a combination thereof, but these do not control the disease’s recurrence. Among the various nanomaterials discussed, liposomes and polymeric nanoformulations have demonstrated encouraging outcomes for facilitated brain cancer treatment. Conclusions: Nanomaterials possess the capacity to overcome the shortcomings of conventional therapies. Polymer-based nanomaterials have shown encouraging outcomes against brain cancer when amalgamated with other nano-based therapies. Nonetheless, nanomaterials could be devised that possess minimal toxicity towards normal cells or that specifically target tumor cells. In addition, rigorous clinical investigations are warranted to prepare them as an efficient and safe modality for brain cancer therapy.

Cyanobacteria-From the Oceans to the Potential Biotechnological and Biomedical Applications.

Cyanobacteria are photosynthetic prokaryotic organisms which represent a significant source of novel, bioactive, secondary metabolites, and they are also considered an abundant source of bioactive compounds/drugs, such as dolastatin, cryptophycin 1, curacin toyocamycin, phytoalexin, cyanovirin-N and phycocyanin. Some of these compounds have displayed promising results in successful Phase I, II, III and IV clinical trials. Additionally, the cyanobacterial compounds applied to medical research have demonstrated an exciting future with great potential to be developed into new medicines. Most of these compounds have exhibited strong pharmacological activities, including neurotoxicity, cytotoxicity and antiviral activity against HCMV, HSV-1, HHV-6 and HIV-1, so these metabolites could be promising candidates for COVID-19 treatment. Therefore, the effective large-scale production of natural marine products through synthesis is important for resolving the existing issues associated with chemical isolation, including small yields, and may be necessary to better investigate their biological activities. Herein, we highlight the total synthesized and stereochemical determinations of the cyanobacterial bioactive compounds. Furthermore, this review primarily focuses on the biotechnological applications of cyanobacteria, including applications as cosmetics, food supplements, and the nanobiotechnological applications of cyanobacterial bioactive compounds in potential medicinal applications for various human diseases are discussed.

Green synthesis of lanthanum oxide nanoparticles using Moringa oleifera leaves extract and its biological activities

Phytosynthesis is a reliable way to produce metal nanoparticles without affecting the environment. Plant extracts act as reducing agent and favors nanoparticle synthesis. Recently, potential drugs were developed in nanotechnology platforms by the green synthesis approach. In this study, the leaves extract of ‘Moringa Oleifera’ (M. oleifera) used as a reducing agent for the synthesis of Lanthanum oxide nanoparticles (La2O3 NPs). The X-ray diffraction (XRD) confirmed the formation of body-centered cubic structure of La2O3 NPs. The optical behavior of La2O3 NPs was analyzed by UV–Vis spectrum. The bandgap energy of the La2O3 NPs was found to be 4.31 eV using Tauc’s plot. The morphology and purity of La2O3 NPs was analyzed by using Field Emission Scanning Electron Microscope (FESEM) and Energy Dispersive X-ray (EDX) spectrum. High Resolution Transmission Electron Microscope (HR-TEM) analysis reveals the morphology, lattice spacing, and selected area electron diffraction (SAED) pattern of the La2O3 NPs. The XPS analysis of the La2O3 NPs reveals the binding energy of La (3d5/2 and 3d3/2) and O 1s at 835.5, 852.3, and 536 eV respectively. The total antioxidant activity (TOA) of La2O3 NPs was found to be 75.32% at 500 µg/mL with the standard drug of vitamin C. The anti-inflammatory activity of the La2O3NPs was found to be 94.15% at 500 µg/mL using the bovine serum albumin denaturation (BSA) technique. The inhibitory activity of La2O3 NPs against α-amylase was found to be 79.99% at 500 µg/mL. In summary, the pure, highly stable and good biocompatible, greener approach based M. oleifera assisted La2O3 was synthesized for radical scavenging, α-amylase and BSA denaturation inhibition activities which can play a key role in the future biomedical and nano-biotechnological applications.

Factors deciding the assembly and thermostability of the DmrB cage

Dihydromethanopterin reductase (DmrB), is a naturally occurring cage protein found in various archaeal and a few bacterial species. It exists as 24mer with cubic geometry where 8 trimeric subunits are present at the corners of each cube. Each trimer is made up of three monomeric units and six FMN, where two molecules of FMN are present at the interface of each monomer. DmrB is involved in the conversion of dihydromethanopterin to tetrahydromethanopterin using FMN as a redox equivalent. In the present study, we have used spectroscopic and biochemical techniques along with complementary bio-informatic work to understand the assembly principles of the DmrB. Our results show a concentration dependant self-assembly of DmrB which is mediated by ionic interactions. The co-factor FMN stabilizes and preserves the secondary and quaternary structure of DmrB against thermal insult, indicating that the higher order assembly of DmrB is very thermostable. Our work provides an interesting piece of information regarding the role of the co-factors in the thermostability of these classes of cage proteins. The understanding of the assembly and disassembly of this thermostable cage would enable the downstream usage of this system in various nano-biotechnological applications.

Induction of Axonal Outgrowth in Mouse Hippocampal Neurons via Bacterial Magnetosomes.

Magnetosomes are membrane-enclosed iron oxide crystals biosynthesized by magnetotactic bacteria. As the biomineralization of bacterial magnetosomes can be genetically controlled, they have become promising nanomaterials for bionanotechnological applications. In the present paper, we explore a novel application of magnetosomes as nanotool for manipulating axonal outgrowth via stretch-growth (SG). SG refers to the process of stimulation of axonal outgrowth through the application of mechanical forces. Thanks to their superior magnetic properties, magnetosomes have been used to magnetize mouse hippocampal neurons in order to stretch axons under the application of magnetic fields. We found that magnetosomes are avidly internalized by cells. They adhere to the cell membrane, are quickly internalized, and slowly degrade after a few days from the internalization process. Our data show that bacterial magnetosomes are more efficient than synthetic iron oxide nanoparticles in stimulating axonal outgrowth via SG.

Nanomaterial-aided seed regeneration in the global warming scenario: multiwalled carbon nanotubes, gold nanoparticles and heat-aged maize seeds

Seeds in soil seed-banks count amongst organisms that are the most vulnerable to the extreme temperatures predicted in certain models of global warming. This work aims to examine the possibility of a nanomaterial-aided resuscitation of heat-damaged hybrid maize (Zea mays L. “Albatross”) seeds through a study of their germinability and growth after high heat exposures. Heat energy at constant ‘dose’ was delivered at the three temperatures of 40, 50 and 60 °C (40–60) and at a much higher dose at 60 °C (+). Seed germinability and seedling growth indices were studied in the presence/absence of multiwalled carbon nanotubes (MWCNT) and our laboratory synthesised and characterised (SEM, DLS, FTIR, UV–Vis) gold nanoparticles (AuNP). Overall, the MWCNT outperformed the AuNP. The seemingly erratic germination outcomes were rationalised from the standpoint of the biomechanics of water entry into seeds with heat-damaged seedcoats. Ficksian diffusion calculations applied to the interstitial channels of the seedcoat and cellwall showed that the former but not the latter, was an effective barrier to MWCNT entry and that both easily allowed water entry. The MWCNT treatments appear to catalyse the recovery of the (+) seeds and seedlings, promote biomass and the root-to-shoot water transport, whilst the parabolic biomass trend for 40–60 suggests opposing temperature-regulated factors possibly involving reactive oxygen species. Summarising, growth indices of heat-damaged seeds show a strong non-monotonic dose dependence where the MWCNT significantly altered the response, catalysing the recovery of the high heat ‘dose’ set. These results hold promise for the application of nano-biotechnology to agriculture in the global warming scenario.

Applications of Nanobiotechnology in Plant Sciences

Nanotechnology is regarded as the science of manipulation of atomic and subatomic particles for developing new machinery....

Biotechnology and Nanotechnology Drug Delivery: A Review

Nanobiotechnology is a blend of biotechnology and nanotechnology. Nanotechnology, as a field, has been tapped into in recent years, in search of alternatives to overcome challenges associated with drug delivery. Various techniques have been put together in order to fabricate nanoparticles with improved therapeutic efficacy enhanced drug stability, specific targeting of the cells and controlled release inside the cell. This review tends to look at the concepts involved in nanobiotechnology, the application of nanobiotechnology in nanomedicine, diagnosis, cancer treatment and drug delivery approaches.