Polydopamine-functionalized polyethersulfone membrane: A paradigm advancement in the field of α-amylase stability and immobilization.

Biocatalytic membranes have great potential in various industrial sectors, with the immobilization of enzymes being a crucial stage. Immobilizing enzymes through covalent bonds is a complex and time-consuming process for large-scale applications. Polydopamine (PDA) offers a more sustainable and eco-friendly alternative for enzyme immobilization. Therefore, surface modification with polydopamine as mussel-inspired antifouling coatings has increased resistance to fouling. In this study, α-amylase enzyme was covalently bound to a bioactive PDA-coated polyethersulfone (PES) membrane surface using cyanuric chloride as a linker. The optimal activity of α-amylase enzyme immobilized on PES/PDA membrane was obtained at temperature and pH of 55°C and 6.5, respectively. The immobilized enzyme can be reused up to five reaction cycles with 55 % retention of initial activity. Besides, it maintained 60 % of its activity after being stored for five weeks at 4°C. Additionally, the immobilized enzyme demonstrated increased Michaelis constant and maximum velocity values during starch hydrolysis. The results of the biofouling experiment of various membranes in a dead-end cell demonstrated that the PES membrane's water flux increased from 6722.7 Lmh to 7560.2 Lmh after PDA modification. Although α-amylase immobilization reduced the flux to 7458.5 Lmh due to enhanced hydrophilicity, compared to unmodified membrane. The findings of this study demonstrated that the membrane produced through co-deposition exhibited superior hydrophilicity, enhanced coating stability, and strong antifouling properties, positioning it as a promising candidate for industrial applications.

Bioinspired amino acid-functionalized cobalt ferrite nanocomposite: A nanozyme-based colorimetric sensor for sensitive and selective quantification of phenolic compounds and ascorbic acid antioxidant capacity.

The escalating oxidative stress has heightened the daily human demand for diverse antioxidants. Therefore, development of the novel approaches to assess the total antioxidant capacity (TAC) of various nutrients is essential. In this study, drawing inspiration from the active site of native peroxidase enzymes, a novel peroxidase (POD)-like nanozyme was developed based on the cobalt ferrite (CoFe2O4) nanoparticles functionalized with different catalytic amino acids. Based on the TMB/H2O2 colorimetric system, the most substantial enhancement in POD-like activity was obtained by the glutamic acid coating among different charged amino acids studied, with more than 74% increase in specific activity compared to the bare CoFe2O4. A signal-off colorimetric sensing platform based on the obtained nanobiocatalyst was developed for the accurate quantification of the antioxidant capacity of phenolic compounds and vitamin C. The sensitive and selective quantification of ascorbic acid, tannic acid, gallic acid, cyanidin-3-glucoside, and quercetin was obtained by this colorimetric method.

Assessment of the efficiency and stability of enzymatic membrane reaction utilizing lipase covalently immobilized on a functionalized hybrid membrane.

Enzyme immobilization in membrane bioreactors has been considered as a practical approach to enhance the stability, reusability, and efficiency of enzymes. In this particular study, a new type of hybrid membrane reactor was created through the phase inversion method, utilizing hybrid of graphene oxide nanosheets (GON) and polyether sulfone (PES) in order to covalently immobilize the Candida rugosa lipase (CRL). The surface of hybrid membrane was initially modified by (3-Aminopropyl) triethoxysilane (APTES), before the use of glutaraldehyde (GLU), as a linker, through the imine bonds. The resulted enzymatic hybrid membrane reactors (EHMRs) were then thoroughly analyzed by using field-emission scanning electron microscopy (FE-SEM), contact angle goniometry, surface free energy analysis, X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, attenuated total reflection (ATR), and energy-dispersive X-ray (EDX) spectroscopy. The study also looked into the impact of factors such as initial CRL concentration, storage conditions, and immobilization time on the EHMR's performance and activity, which were subsequently optimized. The results demonstrated that the CRLs covalently immobilized on the EHMRs displayed enhanced pH and thermal stability compared to those physically immobilized or free. These covalently immobilized CRLs could maintain over 60% of their activity even after 6 reaction cycles spanning 50 days. EHMRs are valuable biocatalysts in developing various industrial, environmental, and analytical processes.

Lysozyme-immobilized bandage contact lens inhibits the growth and biofilm formation of common eye pathogens in vitro.

Bandage contact lenses have an increased affinity to accumulate tear film proteins and bacteria during wear. Among the wide variety of tear film proteins, lysozyme has attracted the most attention for several reasons, including the fact that it is found at a high concentration in the tear film, has exceptional antibacterial and antibiofilm properties, and its significant deposits onto contact lenses. This study aims to evaluate the effect of lysozyme on bacterial biofilm formation on bandage contact lenses. For this purpose, several methods, including microtiter plate test and Colony Forming Unit (CFU) assay have been used to determine antibacterial and antibiofilm characteristics of lysozyme against the two most frequent contact lens-induced bacterial ocular infections, Staphylococcus aureus, and Pseudomonas aeruginosa. The results of these assays demonstrate lysozyme potential to inhibit 57.9% and 80.7% of the growth of S. aureus and P. aeruginosa, respectively. In addition, biofilm formations of P. aeruginosa and S. aureus reduced by 38.3% and 62.7%, respectively due to the antibiofilm effect of lysozyme. SEM and AFM imaging were utilized to visualize lysozyme antibacterial activity and topography changes of the contact lens surface, respectively, in the presence/absence of lysozyme. The results indicated that lysozyme can efficiently attack both gram-positive and gram-negative bacteria and consequently lysozyme-functionalized bandage contact lenses can reduce the risk of ocular infection after eye surgery.

Functionalized graphene oxide/Fe3O4 nanocomposite: A biocompatible and robust nanocarrier for targeted delivery and release of anticancer agents.

The development of efficient drug nanocarriers has remained an important challenge in advanced drug delivery in human body. Combination of graphene-based nanomaterials and cyanuric chloride (CC), as a linker, may improve the success of drug delivery. Herein, a simple approach was used for the synthesis of superparamagnetic graphene oxide (SPMGO) nanocomposite through a chemical precipitation method. The nanocomposite was readily functionalized with cyanuric chloride as a linker for loading the drug. The FTIR spectroscopy confirmed the efficient synthesis of nanocarriers. So did the transmission electron microscopy, atomic force microscopy, and thermo-gravimetric analysis, X-ray diffraction and X-ray photoelectron spectroscopy. Subsequently, the synthesized nanocarriers were studied in terms of their potential for biomedical applications. Immobilization of methotrexate (MTX), as a drug for treatment of cancer was taken into action on the SPMGO and SPMGO/CC. The in vitro assays indicated that the drug nanocarrier systems, SPMGO/MTX and SPMGO/CC/MTX, are hemo-compatible and increase the efficiency of MTX against Caov-4, HeLa and MCF-7 cell lines. The MTX nanocarriers represented a considerably high drug loading and controlled drug release. The overall results indicated the great potential of SPMGO/CC/MTX nanocarrier for targeted drug delivery, particularly in MTX chemotherapy.

Isolation of HLA-G+ cells using MEM-G/9 antibody-conjugated magnetic nanoparticles for prenatal screening: a reliable, fast and efficient method.

The development of an effective and noninvasive early method for obtaining fetal cells is crucial to prenatal screening. Despite proving the presence of fetal cells in the reproductive tract, their use is limited due to their inability to properly isolate them from maternal cells. Magnetic-activated cell sorting (MACS) is a simple technique to separate cells. The present study aimed to develop a MACS-based platform for the isolation of the HLA-G expressing trophoblast cells. For this purpose, first, the triazine functionalized MNPs were synthesized and characterized. Then, MNPs were directly and indirectly conjugated by the MEM-G/9 antibodies targeting HLA-G+ cells. The antibody amount on the surface of the nanoparticles was determined with the Bradford assay. The cell capture efficiency was also investigated. Various characterization methods confirmed the successful nanoparticle synthesis and antibody conjugation. The optimal initial antibody amount for the immobilization was about 20 µg and the optimal time was 3 h. The antibody-nanoparticles by the indirect method had better targeting and capture efficiency than the direct method. The MNPs indirectly conjugated with antibodies are an efficient tool for cell isolation and present considerable potential to be applied in biomedical fields.

Recent developments in enzyme immobilization technology for high-throughput processing in food industries.

The demand for food and beverage markets has increased as a result of population increase and in view of health awareness. The quality of products from food processing industry has to be improved economically by incorporating greener methodologies that enhances the safety and shelf life via the enzymes application while maintaining the essential nutritional qualities. The utilization of enzymes is rendered more favorable in industrial practices via the modification of their characteristics as attested by studies on enzyme immobilization pertaining to different stages of food and beverage processing; these studies have enhanced the catalytic activity, stability of enzymes and lowered the overall cost. However, the harsh conditions of industrial processes continue to increase the propensity of enzyme destabilization thus shortening their industrial lifespan namely enzyme leaching, recoverability, uncontrollable orientation and the lack of a general procedure. Innovative studies have strived to provide new tools and materials for the development of systems offering new possibilities for industrial applications of enzymes. Herein, an effort has been made to present up-to-date developments on enzyme immobilization and current challenges in the food and beverage industries in terms of enhancing the enzyme stability.

Assessment of a New Ginsenoside Rh2 Nanoniosomal Formulation for Enhanced Antitumor Efficacy on Prostate Cancer: An in vitro Study.

INTRODUCTION: Ginsenoside Rh2, purified from the Panax ginseng root, has been demonstrated to possess anticancer properties against various cancerous cells including colorectal, breast, skin, ovarian, prostate, and liver cancerous cells. However, the poor bioavailability, low stability on gastrointestinal systems, and fast plasma elimination limit further clinical applications of Ginsenoside Rh2 for cancer treatments. In this study, a novel formulation of niosomal Ginsenoside Rh2 was prepared using the thin film hydration technique. METHODS: The niosomal formulation contained Span 60 and cholesterol, and cationic lipid DOTAP was evaluated by determining particle size distribution, encapsulation efficiency, the polydispersity index (PDI), and surface morphology. The cytotoxic effects of free Ginsenoside Rh2 and Ginsenoside Rh2-loaded niosomes were determined using the MTT method in the PC3 prostate cancer cell line. For the investigation of the in vitro cellular uptake of Ginsenoside Rh2-loaded niosome, two formulations were prepared: the Ginsenoside Rh2-loaded niosomal formula containing 5% DOTAP and the Ginsenoside Rh2-loaded niosomal formula without DOTAP. RESULTS: The mean size, DPI, zeta potential, and encapsulation efficiency of the Ginsenoside Rh2-loaded nanoniosomal formulation containing DOTAP were 93.5±2.1 nm, 0.203±0.01, +4.65±0.65, and 98.32% ±2.4, respectively. The niosomal vesicles were found to be round and have a smooth surface. The release profile of Ginsenoside Rh2 from niosome was biphasic. Furthermore, a two-fold reduction in the Ginsenoside Rh2 concentration was measured when Ginsenoside Rh2 was administered in a nanoniosomal form compared to free Ginsenoside Rh2 solutions in the PC3 prostate cancer cell line. After storage for 90 days, the encapsulation efficiency, vesicle size, PDI, and zeta potential of the optimized formulation did not significantly change compared to the freshly prepared samples. The cellular uptake experiments of the niosomal formulation demonstrated that by adding DOTAP to the niosomal formulation, the cellular uptake was enhanced. DISCUSSION: The enhanced cellular uptake and cytotoxic activity of the Ginsenoside Rh2 nanoniosomal formulation on the PC3 cell make it an attractive candidate for application as a nano-sized delivery vehicle to transfer Ginsenoside Rh2 to cancer cells.

Xylanase immobilization onto trichlorotriazine-functionalized polyethylene glycol grafted magnetic nanoparticles: A thermostable and robust nanobiocatalyst for fruit juice clarification.

The covalent immobilization of xylanase onto the trichlorotriazine-functionalized polyethylene glycol grafted magnetic nanoparticles was exploited to generate a stabilized xylanase with improved catalytic activity and stability. Several tools were deployed to monitor the synthesis and immobilization processes, the loading capacity of nanocarrier, and the structural/chemical characteristics of the nanobiocatalyst. The optimum immobilization yield of xylanase was 260 mg xylanase/g nanocarrier in 20 mM phosphate buffer, pH 6.5 at 25 °C. A forward shift in optimum pH (6.5 to 7.5) and temperature (60 to 70 °C) of xylanase was observed after immobilization and the performance of immobilized enzyme was improved at high temperatures and pHs as affirmed by enhancement of vmax (2.69 to 6.01 U/mL) and decreases of Ea (14.61 to 13.41 kJ/mol). An increase in Km from 25.51 to 40.42 mg/mL was recorded after immobilization. The obtained results indicated augmented thermal stability of the immobilized xylanase. Notably, it showed good reusability as validated by retention of 50% of its initial activity after nine recycles in enrichment of the pineapple juice clarification after 120 min incubation at 50 °C, pH 4.5. The structural analysis revealed some partial changes in the α-helix and ß-sheet content of the enzyme after several recycles.

The correlation of combined OGG1, CYP1A1 and GSTP1 gene variants and risk of lung cancer of male Iraqi waterpipe tobacco smokers.

Genetic polymorphisms of genes whose products are responsible for activities, such as xenobiotic metabolism, mutagen detoxification and DNA-repair, have been predicted to be associated with the risk of developing lung cancer (LC). The association of LC with tobacco smoking has been extensively investigated, but no studies have focused on the Arab ethnicity. Previously, we examined the association between genetic polymorphisms among Phase I and Phase II metabolism genes and the risk of LC. Here, we extend the data by examining the correlation of OGG1 Ser326Cys combined with CYP1A1 (Ile462Val and MspI) and GSTP1 (Ile105Val and Ala103Val) polymorphisms with the risk of LC. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and gene sequencing were carried out for genotyping the OGG1 polymorphisms of 123 LC patients and 129 controls. No significant differences in the frequencies of the OGG1 mutant allele between patients and controls were found. The distributions of heterozygous Ser/Cys or Cys/Cys genotypes of OGG1 were not associated with the risk of LC either according to the histological types of LC or based on waterpipe tobacco (WP) smoking status. In contrast, the combined effect of OGG1 variants with CYP1A1 and GSTP1 variants revealed a significant correlation with the OGG1 Ser326Cys-CYP1A1 MspI variants pairing. This association was significant (p = 0.001) in individuals who carried homozygous or heterozygous variant type genotypes of both genes in a reference with carriers of both wild-type genotypes (wt/wt - wt/wt). The odds ratios were 2.99 (95% CI 1.67-5.36), 2.68 (95% CI 1.08-6.62), and 2.80 (95% CI 1.18-6.69) for those who carried (wt/wt - wt/vt + vt/vt), (wt/vt + vt/vt - wt/wt), and (wt/vt + vt/vt - wt/vt + vt/vt), respectively. The study suggests a limited correlation is present between carrying OGG1 Ser326Cys polymorphism and the risk of developing LC in Arab populations.

Efficient immobilization of pectinase on trichlorotriazine-functionalized polyethylene glycol-grafted magnetic nanoparticles: A stable and robust nanobiocatalyst for fruit juice clarification.

Developing an effective strategy to economically exploitation of pectinase, as one of the most widely used enzymes in food industry, is of utmost importance. Herein, pectinase was covalently immobilized onto polyethylene glycol grafted magnetic nanoparticles via trichlorotriazine with high loading efficiency. The generated immobilized pectinase showed enhanced catalytic activity, improved operational stability, and easily reusability. Thermal and pH stabilities studies showed improved performance of immobilized pectinase especially at extreme points. Compared to free enzyme, the noticeably lower Km and higher vmax values of immobilized pectinase demonstrated the enhanced catalytic activity of this enzyme after immobilization. Besides, the immobilized enzyme exhibited excellent reusability and stability by retaining up to 55 and 94% of its initial activity after 10 recycles and 125 days storage at 25 °C, respectively. Moreover, turbidity reduction occurred up to 59% in treated pineapple juice with immobilized pectinase, suggesting applicability of this system in juice and food-processing industries.

Correlation of GSTP1 gene variants of male Iraqi waterpipe (Hookah) tobacco smokers and the risk of lung cancer.

Glutathione-S-transferases (GSTs) play a role in the detoxification of environmental chemicals and mutagens, such as those inhaled during tobacco smoking. There have been conflicting reports concerning GST polymorphisms as risk factors in the development of lung cancer. No studies focused on Arab populations exposed to Waterpipe (WP) tobacco smoke have been undertaken. Here Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and gene sequencing were applied to analyze allelic variations in GSTP1-rs1695 and -rs1138272 amongst 123 lung cancer patients and 129 controls. The data suggest that WP smoking raised the risk of lung cancer more than three-fold (OR 3.6; 95% CI 2.1-6.0; p < 0.0001). However, there was no significant association between individual GSTP1 polymorphisms and the risk of lung cancer. In contrast, analysis of the rs1695 and rs1138272 combination suggested that the risk of lung cancer was raised more than two-fold for carriers of the GSTP1-rs1695 (G) allele (OR 2.5; 95% CI 1.0-6.4; p < 0.05), however, the presence of the GSTP1-rs1138272 (T) allele, in addition to GSTP1-rs1695, did not significantly change the risk ratio (OR 2.8; 95% CI 1.4-5.7; p < 0.004). WP tobacco smokers who carried the GSTP1-rs1695, but not GSTP1-rs1138272, allele were similarly susceptible to lung cancer (OR 2.4; 95% CI 1.1-5.3; p < 0.03). Hence, the results suggest that smoking WP tobacco and carrying GSTP1-rs1695 polymorphisms are risk factors for lung cancer in Arab Iraqi males.

Surface modification of polypropylene membrane for the removal of iodine using polydopamine chemistry.

The development of stable and effective iodine removal systems would be highly desirable in addressing environmental issues relevant to water contamination. In the present research, a novel iodine adsorbent was synthesized by self-polymerization of dopamine (PDA) onto inert polypropylene (PP) membrane. This PP/PDA membrane was thoroughly characterized and its susrface propeties was analyzed by various analytical techniques indcluding field emission scanning electron microscopy (FESEM), atomic force microscopy (AFM), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halenda (BJH), contact angle, and surface free energy measurement. The PP/PDA membranes were subsequently used for batchwise removal of iodine at different temperatures (25-70 °C), pH (2-7), and surface areas (1-10 cm2) to understand the underlying adsorption phenomena and to estimate the membrane capacity for iodine uptake. The increase in temperature and pH both led to higher adsorption of iodine. The present approach showed a removal efficiency of over 75% for iodine using 10 cm2 PP/PDA membrane (18.87 m2 g-1) within 2 h at moderate temperatures (∼50 °C) and pH > 4, about 15 fold compared to the PP control membrane. The adsorption kinetics and isotherms were well fitted to the pseudo-second-order kinetic and Langmuir isotherm models (R2 > 0.99). This adsorbent can be recycled and reused at least six times with stable iodine adsorption. These findings were attributed to the homogenous monolayer adsorption of the iodide on the surface due to the presence of catechol and amine groups in the PP/PDA membrane. This study proposes an efficient adsorbent for iodine removal.

Magnetic chitosan-copper nanocomposite: A plant assembled catalyst for the synthesis of amino- and N-sulfonyl tetrazoles in eco-friendly media.

A greener, cost efficient and simple method is described to prepare copper nanoparticles (NPs) immobilized on the magnetic chitosan (one of the more versatile polysaccharides) using Euphorbia falcata leaf extract as reducing/stabilizing agent. The prepared catalyst (Cu NPs@Fe3O4-chitosan) was authenticated by field emission scanning electron microscope (FESEM), X-ray diffraction (XRD), transmission electron microscopy (TEM), Scanning transmission electron microscopy (STEM), energy-dispersive X-ray spectroscopy (EDS), thermogravimetry/derivative thermogravimetry (TG/DTG), Vibrating sample magnetometer (VSM), and elemental mapping. TEM analysis indicates that Cu NPs with average sizes in 5-10 nm range is formed on magnetic chitosan with the spherical morphology. The Cu NPs@Fe3O4-chitosan was employed as a new catalyst for the synthesis of different tetrazoles by the reaction of various secondary or tertiary cyanamides with sodium azide in water under reflux conditions. Easy separation by external magnetic field, mild reaction conditions, low cost and the reusability are some of the beneficial features of this catalyst.

Template-oriented synthesis of hydroxyapatite nanoplates for 3D bone printing.

The design of hydroxyapatite (HA) nanoarchitecture is critical for fabricating artificial bone tissues as it dictates the biochemical and the mechanical properties of the final product. Herein, we incorporated a simple hard-template approach to synthesise single crystal nanoplates of HA. We used the 2D graphitic nitride (g-C3N4) material to prepare an HA sol-gel under hydrothermal conditions. A new HA nanostructure was then formed during the calcination and removal of g-C3N4 at a higher temperature, which finally led to the production of nanoplates (thickness of ∼100 nm) while in lateral dimension the average size was in the micrometre scale. We characterised the synthesised HA nanoplates with XRD, TEM, and HRTEM. The theoretically predicted nanostructure construction based on Wulff's method is in full agreement with the experimental observations. We then prepared different weight ratios of HA and polylactic acid (PLA) composites for artificial 3D bone fabrication. The strong interaction between PLA and HA's (110) facet, which was the second most prevalent, resulted in the composite's mechanical robustness. After mechanical testing, an optimum ratio was selected for biological studies and 3D printing. Biological experiments demonstrated that the synthesised composite had excellent viability in vitro.

PEGAylated graphene oxide/superparamagnetic nanocomposite as a high-efficiency loading nanocarrier for controlled delivery of methotrexate.

Polymer-coated nanocarriers play an important role in targeted drug delivery. The use of polymers such as polyethylene glycol increases stability, biocompatibility, and blood circulation time of the drug, and may consequently improve the success of drug delivery. In the present work, a simple approach has been reported for synthesizing polyethylene glycol bis amin (PEGA) functionalized graphene oxide/iron oxide nanocomposite as a remarkable unit for loading drugs. The biomedical applications of the synthesized nanocomposite were investigated by immobilizing methotrexate (MTX), as an anticancer drug. The structural and morphological characteristics and the successful synthesis of the nanocomposite were evaluated by different charachterization techniques. The cytotoxicity assay of the nanocarrier showed higher toxicity against HeLa and MCF-7 cell lines, compared to free MTX. The drug release experiments in acidic and physiological conditions suggested the first order kinetics model for the release of MTX from the nanocomposite. Furthermore, the agglutination, complement activation, and coagulation time experiments demonstrated the blood compatibility of the synthesized nanocarrier.

Hybrid and Composite Scaffolds Based on Extracellular Matrices for Cartilage Tissue Engineering.

IMPACT STATEMENT: Scaffolds fabricated from extracellular matrix (ECM) derivatives are composed of conducive structures for cell attachment, proliferation, and differentiation, but generally do not have proper mechanical properties and load-bearing capacity. In contrast, scaffolds based on synthetic biomaterials demonstrate appropriate mechanical strength, but the absence of desirable biological properties is one of their main disadvantages. To integrate mechanical strength and biological cues, these ECM derivatives can be conjugated with synthetic biomaterials. Hence, hybrid scaffolds comprising both advantages of synthetic polymers and ECM derivatives can be considered a robust vehicle for tissue engineering applications.

Preparation of a stable and robust nanobiocatalyst by efficiently immobilizing of pectinase onto cyanuric chloride-functionalized chitosan grafted magnetic nanoparticles.

In industrial processes, effective degradation of polygalacturonic acid using immobilized pectinase is preferred over free one due to its stability and efficient functional reuses. Pectinase was covalently conjugated to the surface of cyanuric chloride functionalized chitosan encapsulated magnetite nanoparticles. The results obtained of various analytical tools and biochemical studies demonstrated successful synthesis and immobilization processes, high immobilization efficiency and loading capacity. The circular dichroism (CD) results of free and immobilized pectinase revealed the partial decreases in the α-helices and ß-sheets, and marginal increases in the unordered elements contents of pectinase upon the immobilization onto Fe3O4@Ch-CC nanoparticles, along with stability improvement. The immobilized pectinase was retained about 60% of its initial catalytic activity after 13 recycles at optimum conditions (40 °C, pH 4.5). The storage stability of pectinase was increased due to immobilization, after 75 days storage at 4 °C, the free and immobilized enzyme retained 43% and 74% of the initial activity, respectively. The immobilized pectinase showed higher storage stability and better performance at wider ranges of pH and temperature, compared to free pectinase.

Enhancement of Tryptic Digestibility of Milk ß-Lactoglobulin Through Treatment with Recombinant Rice Glutathione/Thioredoxin and NADPH Thioredoxin Reductase/Thioredoxin Systems.

ß-Lactoglobulin (BLG), a member of lipocalin family, is one of the major bovine milk allergens. This protein exists as a dimer of two identical subunits and contains two intramolecular disulfide bonds that are responsible for its resistance to trypsin digestion and allergenicity. This study aimed to evaluate the effect of reduction of disulfide bonds of BLG with different rice thioredoxins (Trxs) on its digestibility and allergenicity. Therefore, the active recombinant forms of three rice Trx isoforms (OsTrx1, OsTrx20, and OsTrx23) and one rice NADPH-dependent Trx reductase isoform (OsNTRB) were expressed in Escherichia coli. Based on SDS-PAGE, HPLC analysis, and competitive ELISA, the reduction of disulfide bonds of BLG with OsNTRB/OsTrx23, OsNTRB/OsTrx1, GSH/OsTrx1, or GSH/OsTrx20 increased its trypsin digestibility and reduced its immunoreactivity. The finding of this study opens new insights for application of plant Trxs in the improvement of food protein digestibility. Especially, the use of OsTrx20 and OsTrx1 are more cost-effective than E. coli and animal Trxs due to their reduction by GSH and no need to NADPH and Trx reductase as mediator enzyme.

Enzyme Immobilization on Functionalized Graphene Oxide Nanosheets: Efficient and Robust Biocatalysts.

Enzymes are used as biocatalysts for analytical purposes in diagnostics and preparative purposes in large-scale industrial processes. Despite perfect catalytic properties of enzymes, their industrial applications are limited due to the drawbacks regarding the lack of long-term stability under process conditions. The difficulties associated with recycling have to be resolved before enzyme implementation at industrial scale. Enzyme immobilization, as a novel approach, can improve the half-life, stability, catalytic activity, and reusability of enzymes. Graphene-based nanomaterials, as nanoscaled and thermostable inorganic carriers, are nontoxic materials and selective modulators for enzyme activity. Herein, we have concentrated on strategies for preparing graphene-based nanocomposites for enzyme immobilization. Nanostructures of graphene, hybrid graphene, and their derivatives with adjustable surface chemistry, caused them to be excellent candidates for immobilization of enzymes. For instance, the synthesis and functionalization of Fe3O4-graphene oxide (GO) hybrids were improved recently, in our research group, using cyanuric chloride and polyethylene glycol bis-amine for the immobilization of xylanase and glucoamylase enzymes, via physical and covalent attachments. Decorating GO nanosheets with Fe3O4 nanoparticles has facilitated the reusability of enzymes and increased the surface area for enzyme loading. The use of these hydrophilic crosslinkers may change the microenvironment of the immobilized enzymes that could result in the enhancement of their catalytic activity. As a result of the fascinating properties of graphene-based nanocarriers, with respect to structures that can be oriented and surfaces that can be modified, in our opinion, they offer some important advantages for biotechnological applications, especially in the areas of enzyme immobilization and medicine.