Immobilization Efficiency Research Articles

Performance and hydration mechanism of MSWI FA-barium slag-based all-solid waste binder

Barium slag (BS) and municipal solid waste incineration fly ash (MSWI FA) are hazardous wastes produced in large quantities, with heavy metals such as Ba, Pb, and Cr significantly exceeding acceptable levels, posing a threat to the ecological environment and living organisms. The development of technologies for the harmless disposal and resource utilization of hazardous waste is a current research focus. This study used BS, blast furnace slag (BFS), MSWI FA, and flue gas desulphurized (FGD) gypsum to replace cement in the preparation of solid waste-based binder. The results indicated that the optimal mass ratio was BS: BFS: FGD gypsum: MSWI FA = 14%:56%:20%:10%. The compressive strengths of the binder at 3, 7,and 28 days were 23.7MPa, 30.76MPa, and 38.65MPa, respectively. Chinese standard HJ557-2010 was adopted to carry out heavy metal leaching experiment on raw materials, the immobilization efficiency of heavy metals Ba, Pb, and Cr were 99%, 99%, and 96%, respectively, and the leachate toxicity was below the Class III groundwater limit. XRD, FT-IR, TG-DSC, XPS, SEM-EDS and other experimental methods were used to study the effect of BS content on the properties of cementable materials. An appropriate amount of BS hydrolyzes during hydration, releasing Ba²⁺ and OH⁻ ions, which activated the BFS, producing AFt and C–(A)–S–H gel. These products interweaved with material particles to fill the pore spaces, enhancing the binder’s mechanical strength and heavy metal immobilization capability. However, when excessive BS was added, the hydration products favored the formation of FS, leading to a decline in binder performance.

Biochemical properties of immobilized horseradish peroxidase on ceramic and its application in removal of azo dye.

In the current work, electrostatic interactions were used to immobilize the horseradish peroxidase (HRP) onto five types of ceramic materials (C) with different concentrations of oxidized metals (C1-C5). The highest immobilization efficiency (70 and 77%) was detected at 6mg C3 and 18 enzyme units. Scanning Electron Microscope (SEM), Energy Dispersive X-ray (EDX) and Fourier Transform Infrared (FTIR) analysis of C3-HRP confirmed the immobilization of the enzyme. After ten reuses, the reusability analysis showed that (66%) of the C3-HRP enzyme activity was retained. For C3-HRP, the optimum pH and temperature of the soluble enzyme were shifted from 7.0 and 30°C to 6.0 and 50°C. Up to 40°C and 50°C, respectively, the soluble HRP and C3-HRP remained steady. The kinetic analysis revealed that the Km and Vmax of soluble HRP and C3-HRP were, respectively, 5.5mM, 0.66 units, and 8mM, 0.52 units for hydrogen peroxide (H2O2) and 35.5mM, 3.4 units and 40mM, 1.1 units for guaiacol. Compared to soluble-HRP, the C3-HRP exhibited a greater oxidizing affinity toward several phenolic compounds (Guaiacol, o-dianisidine, o-phenylenediamine, pyrogallol, p-aminoantipyrine). In comparison with soluble-HRP, the C3-HRP showed increased stress tolerance with Triton X-100, urea, metals, isopropanol, and dimethyl sulfoxide. The C3-HRP removed methyl orange more effectively compared to soluble-HRP.

Stability of immobilized L-arginine deiminase from Penicillium chrysogenum and evaluation of its anticancer activity

The aim of the present work was to immobilize L-arginine deiminase on suitable supports such as chitosan, alginate, and silica gel to study its stability. Additionally, the study aims to investigate the anticancer effects of the free purified enzyme on hepatocellular carcinoma (Hep-G2) and breast cancer (MCF-7) cell lines. L-arginine deiminase (ADI: EC 3.5.3.6) was immobilized on chitosan, Ca-alginate, and silica gel, with immobilization efficiencies of 89.0%, 72.8%, and 66.5%, respectively. The optimal immobilization time for the highest efficiency was 4 h. Increasing the concentration of glutaraldehyde improved the immobilization efficiency of ADI on chitosan. The chitosan-immobilized ADI retained about 45% of its activity after 8 cycles. The optimal pH values were 6 for the free purified ADI and 7 for the chitosan-immobilized ADI. The optimal temperature increased from 40 °C for the free enzyme to 45 °C after immobilization. The activation energies for the free and chitosan-immobilized enzymes were 71.335 kJ/mol and 64.011 kJ/mol, respectively. The Km values for the free and chitosan-immobilized ADI were 0.76 mM and 0.77 mM, respectively, while the Vmax values were 80.0 U/mg protein for the free ADI and 71.4 U/mg protein for the chitosan-immobilized ADI. After 30 days of storage at 4 °C, the residual activities were 40% for the free purified ADI and 84% for the chitosan-immobilized ADI. At 25 °C, the residual activities were 10% for the free ADI and 75% for the chitosan-immobilized ADI. The chitosan-immobilized ADI exhibited significantly higher stability against proteases such as pepsin and trypsin compared to the free enzyme. The purified ADI also demonstrated enhanced potential anticancer effects and significant cytotoxicity against the Hep-G2 and MCF-7 tumor cell lines compared to doxorubicin. These findings suggest that purified ADI has potential as an anticancer agent, though further in-depth studies are required.

Endomelanconiopsis endophytica Lipase Immobilized in Calcium Alginate for Production of Biodiesel from Waste Cooking Oil

The increasing global demand for biodiesel is due to the urgent need to replace fossil diesel with a fuel based on renewable energy sources. Although chemical catalysis is widely used to produce biodiesel, it uses harsh operating conditions, has high energy consumption, and generates unwanted byproducts. In this scenario, biocatalysis stands out as an efficient and environmentally friendly alternative to chemical catalysis. In biocatalysis, the use of immobilized enzymes plays an important role in the reduction in costs. In this sense, we investigated the use of the lipase produced by an Amazonian endophytic fungus in an immobilized form in the transesterification of waste cooking oil for biodiesel production. The fungus Endomelanconiopsis endophytica QAT_7AC demonstrated a high production of lipase. The lipolytic extract was precipitated in ethanol, which increased the specific enzyme activity. The lipolytic extract and the precipitated lipolytic extract were immobilized in calcium alginate beads. Immobilization efficiency was over 89%. The immobilized biocatalysts showed thermal stability and were used in the production of biodiesel using waste cooking oil and ethanol. It was possible to reuse them for up to four reaction cycles, with yields greater than 70%. These results prove the efficiency of immobilized biocatalysts in the production of biodiesel from waste oils.

Plasma Deposition of Parylene-like Films with Chemical Functional Groups for Immunoassays.

Parylene-like films obtained via the plasma decomposition of parylene precursors with functional groups (amino and formyl) are proposed as an alternative to those obtained via the thermal method. To analyze the chemical functional groups after plasma deposition, a surface analysis of the parylene films using the two different deposition methods was performed via Fourier transform infrared (FT-IR) and X-ray photoelectron spectroscopy (XPS). The FT-IR analysis revealed that the featured peaks of the chemical functional groups were maintained in the parylene-like films obtained via the plasma deposition method. The XPS analysis revealed that the featured chemical functional groups of parylene-AM and parylene-H were maintained after plasma deposition. The surface energy of the parylene films was estimated by using contact angle measurements. The plasma-deposited parylene films were then employed for protein immobilization via the functional groups using horseradish peroxidase (44 kDa) and green fluorescent protein (25 kDa) as model proteins. The parylene-AM and parylene-H films obtained via plasma deposition exhibited higher immobilization efficiencies than did the same parylene films obtained via thermal deposition. Finally, a competitive immunoassay was obtained by immobilizing the Fv-antibodies on plasma-deposited parylene-AM and parylene-H films via covalent bonding. Using heat-deactivated SARS-CoV-2 as a real sample, the limit of detection at the feasible level for the medical diagnosis of COVID-19 was achieved using a competitive immunoassay based on immobilized Fv-antibodies on plasma-deposited parylene films.

Novel N-doped carbon nanotubes impregnated Mn spheres with polydopamine coating as an efficient polysulfide immobilizer for Li-S batteries

Lithium-sulfur (Li-S) batteries are one of the most promising energy storage and conversion devices due to the high theoretical capacity and cost-effectiveness of sulfur. However, they still suffer from sluggish redox kinetics and the shuttle effect caused by complex polysulfides. In this work, graphitic carbon nitride (g-C3N4) is utilized as a template and further hydrothermally treated with an Mn source and glucose. The pyrolysis of g-C3N4 gives rise to N-doped carbon nanotubes, producing abundant sites for physical confinement and chemical adsorption of polysulfides, while glucose carbonization brings forth amorphous carbon and Mn source produces metal spheres. Afterward, polydopamine (PDA) induces N-doped carbon coating and promotes interface connection as well as electron immigration. This synergistic design possesses a high surface area of micropores and mesopores to aggregate sulfur and accelerate redox kinetics. As a result, the N-doped carbon nanotube with Mn spheres and PDA coating@sulfur (CN/Mn-PDA@S) exhibits a high reversible capacity of 813.5 mAh g−1 at 1 C with a decay rate of 0.064% per cycle and remarkable capacity retention at 2 C with rate performance up to 4 C. Therefore, the novel design of N-doped carbon nanotubes with Mn spheres and PDA coating serves as an efficient polysulfide immobilizer for Li-S batteries.

Improving enzyme immobilization: A new carrier-based magnetic polymer for enhanced covalent binding of laccase enzyme

In the quest for effective enzyme immobilization methods, this study focuses on synthesizing carrier-based magnetic polymer to enhance the covalent binding of T. versicolor laccase. Utilizing chitosan (CS) and alginate (ALG) composites, modified with Fe3O4 magnetic nanoparticles (MNPs), we aimed to improve the enzyme's stability, reusability, and performance under varying conditions. The ionic gelation method was employed to prepare CS-ALG and CS-ALG-Fe3O4 MNPs composites, resulting in an 84 % and 91 % immobilization efficiency. The immobilized enzyme demonstrated superior thermal stability, retaining 48 % activity for CS-ALG-laccase and 67 % for CS-ALG-Fe3O4 MNPs-laccase at 70 °C, compared to 29 % for the free enzyme. Scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy were used to characterize the composites, revealing significant morphological changes and successful enzyme integration. Kinetic studies indicated that immobilization increased the Vmax to 141 μmol/min for CS-ALG-laccase and 111 μmol/min for CS-ALG-Fe3O4 MNPs-laccase, while slightly reducing substrate affinity (Km) to 1.42 mM−1 and 1.32 mM−1, respectively. The immobilized laccase retained higher activity after ten reaction cycles (81 % activity for CS-ALG-Fe3O4 MNPs-laccase) and during prolonged storage (75 % activity retention), showcasing its potential for industrial applications. Additionally, the enzyme exhibited increased resistance to various organic solvents, enhancing its practical utility.

Biochar as an Enzyme Immobilization Support and Its Application for Dye Degradation

Wastewaters generated by the textile industry often contain significant amounts of harmful (carcinogenic and mutagenic) cationic dyes, whose efficient removal is of crucial importance. This study investigates the laccase immobilization on biochar obtained from sour cherry stones (SCS-B), as a cost effective adsorbent, and evaluates its application for brilliant green (BG) degradation. The successful immobilization of laccase on biochar was achieved via adsorption and confirmed using scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX) and Fourier transform infrared spectroscopy (FTIR). An immobilization efficiency of 66% was achieved using 0.274 U/mL of laccase at pH 5 and a temperature of 40 °C. The adsorption kinetics of laccase followed a pseudo-second-order model, indicating that chemical adsorption plays a significant role in the immobilization process. The BG degradation by immobilized system was further optimized by evaluating effects of pH, temperature, dye concentration, and contact time. More than 92% of BG (50 mg/L) was removed within 4 h at pH 5 and temperature of 30 °C. These findings suggest that SCS-B can effectively be used as an enzyme carrier and be further utilized for the removal of emerging pollutants, positioning it as a sustainable solution for wastewater treatment.

Organic and Inorganic Modifications to Increase the Efficiency in Immobilization of Heavy Metal (Zn) in Cementitious Composites-The Impact of Cement Matrix Pore Network Characteristics.

This research investigated the properties of modified cementitious composites including water purification from heavy metal-zinc. A new method for characterizing the immobilization properties of tested modifiers was established. Several additions had their properties investigated: biochar (BC), active carbon (AC), nanoparticulate silica (NS), copper slag (CS), iron slag (EAFIS), crushed hazelnut shells (CHS), and lightweight sintered fly ash aggregate (LSFAA). The impact of modifiers on the mechanical and rheological properties of cementitious composites was also studied. It was found that considered additions had a significantly different influence over the investigated properties. The addition of crushed hazelnut shells, although determined as an effective immobilization modifier, significantly deteriorated the mechanical performance of the composite as well as its rheological properties. Modification by iron slag allowed for a significant increase in immobilization properties (five-fold compared to the reference series) without a substantial impact on other properties. The negative effect on immobilization efficiency was observed for nanoparticulate silica modification due to its sealing effect on the pore network of the cement matrix. The capillary pore content in the cement matrix was identified as a parameter significantly influencing the immobilization potential of most considered modifications, except biochar and active carbon.

Recent Advances in Enzyme‐based Biofuel Cells Using Glucose Fuel: Achieving High Power Output and Enhanced Operational Stability

AbstractAn enzyme‐based biofuel cell (EBFC) is widely regarded as one of the most efficient power sources for bio‐friendly and implantable medical devices, capable of converting electrochemical reactions into electrical currents under physiological conditions. However, despite its potential, the practical and commercial use of EBFCs is limited by their low power output and operational instability. Therefore, significant research efforts have focused on increasing power output and stability by improving electron transfer between enzymes and host electrodes and developing efficient enzyme immobilization techniques. However, most EBFCs produced by current methods still deliver unsatisfactory performance. A promising approach to address these challenges is the use of conductive linkers that promote favorable interfacial interactions between adjacent enzymes and between enzymes and host electrodes. These linkers can facilitate electron transfer and ensure robust enzyme immobilization. In addition, designing the host electrode with a 3D structure and a large surface area can further improve the areal energy performance. This perspective reviews the working principles, types, and electron transfer mechanisms of EBFC electrodes and explores how conductive linkers and 3D host electrodes can enhance the performance of EBFC electrodes. Finally, recent advances in integrating EBFCs into biomedical devices are described.

Ultra-sensitive immobilization-free homogeneous electrochemiluminescence biosensor for thrombin detection via electrostatic interaction and Exo I-powered signal amplification

Herein, we present the development of an ultra-sensitive immobilization-free homogeneous electrochemiluminescence (ECL) biosensor, leveraging the electrostatic repulsion between a negatively charged indium tin oxide (ITO) electrode and DNA and exonuclease I (Exo I)-powered signal amplification, to achieve highly efficient detection of thrombin. Specifically, the aptamer and the complementary DNA engage in the formation of a double-stranded DNA (dsDNA) complex. This negatively charged dsDNA structure subsequently associates with a positively charged ECL indicator, namely ruthenium phenanthroline (Ru(phen)32+), resulting in the generation of the dsDNA-Ru(phen)32+ ECL probe. The negatively charged dsDNA-Ru(phen)32+ experiences electrostatic repulsion from the negatively charged ITO electrode, resulting in a low ECL signal. Nonetheless, upon the addition of thrombin, the aptamer preferentially binds to thrombin, triggering the releases of the embedded Ru(phen)32+ facilitated by Exo I and hence resulting in a robust and enhanced ECL signal. The amplified ECL signal is linearly correlated with the logarithm of thrombin concentration within a detection range spanning from 10 fmol/mL to 50 pmol/mL, with a remarkable detection limit of 3.21 fmol/mL. This strategy eliminates the need for cumbersome labeling steps, avoids the electrode modification process, overcoming the low immobilization efficiency of aptamers and poor signal transduction of indicators labeled at the end of DNA.

Controlled Nanopore Sizes in Supraparticle Supports for Enhanced Propane Dehydrogenation with GaPt SCALMS Catalysts.

The efficient immobilization of GaPt liquid metal alloy droplets onto tailored supports improves catalytic performance by preventing coalescence and subsequent loss of active surface area. Herein, we use tailored supraparticle (SP) supports with controlled nanopores to systematically study the influence of pore sizes on the catalytic stability of GaPt supported catalytically active liquid metal solution (SCALMS) in propane dehydrogenation (PDH). Initially, GaPt droplets were prepared via an atom-efficient and scalable ultrasonication method with recycling loops to yield droplets <300 nm. Subsequently, these droplets were immobilized onto SiO2-based SPs with controlled pore sizes ranging from 45 to 320 nm. Catalytic evaluations in PDH revealed that GaPt immobilized on SPs with larger pores demonstrated superior stability over 15 h time-on-stream evidenced by reduced deactivation rates from 0.046 to 0.026 h-1. Nanocomputed tomography and identical location SEM confirmed the successful immobilization of GaPt droplets within the interstitial sites formed by the primary particles constituting the SPs. These remained unchanged before and after the catalytic reaction, demonstrating efficient coalescence prevention. Our findings underscore the importance of support pore size engineering for improving the stability of GaPt SCALMS catalysts and highlight, particularly, the high potential of using SPs in this context.

Sustainable pretreatment method of lignocellulosic depolymerization for enhanced ruminant productivity using laccase protein immobilized agarose beads

Laccase, the selectively lignin degrader, vital to the initiation of lignocellulosic deconstruction was immobilized onto activated agarose beads to increase its reuse potential. Laccase cross-linked beads (~ 3.42 mm) recorded a specific activity of 23 Umg− 1, retaining about 80.43% enzyme activity after 45 days of storage. The immobilization yield and efficiency were 89% and 97% respectively. The equilibrium data fitted the Freundlich equation (R2 = 0.9987) demonstrating multilayer adsorption and the presence of Cu, Fe, and S in the elemental analysis of immobilized beads established effective binding between activated agarose beads and the laccase protein. Characterization studies of the immobilized laccase-treated crop residues revealed significant differences in the lignin polymer after each treatment cycle. An increase in digestibility of 26.21% and 7.62% was observed in paddy and finger millet-treated straws respectively, over the controls corroborating efficient lignin depolymerization. The propitious performance of laccase beads authenticated in the batch enzymatic reactor to treat crop residues paves headway as a sustainable green technology in the deconstruction of crop residues for use as ruminant feed, augmenting productivity.

Magnetic protein aggregates generated by supramolecular assembly of ferritin cages - a modular strategy for the immobilization of enzymes.

Efficient and cost-effective immobilization methods are crucial for advancing the utilization of enzymes in industrial biocatalysis. To this end, in vivo immobilization methods relying on the completely biological production of immobilizates represent an interesting alternative to conventional carrier-based immobilization methods. This study aimed to introduce a novel immobilization strategy using in vivo-produced magnetic protein aggregates (MPAs). MPA production was achieved by expressing gene fusions of the yellow fluorescent protein variant citrine and ferritin variants, including a magnetically enhanced Escherichia coli ferritin mutant. Cellular production of the gene fusions allows supramolecular assembly of the fusion proteins in vivo, driven by citrine-dependent dimerization of ferritin cages. Magnetic properties were confirmed using neodymium magnets. A bait/prey strategy was used to attach alcohol dehydrogenase (ADH) to the MPAs, creating catalytically active MPAs (CatMPAs). These CatMPAs were purified via magnetic columns or centrifugation. The fusion of the mutant E. coli ferritin to citrine yielded fluorescent, insoluble protein aggregates, which are released upon cell lysis and coalesce into MPAs. MPAs display magnetic properties, as verified by their attraction to neodymium magnets. We further show that these fully in vivo-produced protein aggregates can be magnetically purified without ex vivo iron loading. Using a bait/prey strategy, MPAs were functionalized by attaching alcohol dehydrogenase post-translationally, creating catalytically active magnetic protein aggregates (CatMPAs). These CatMPAs were easily purified from crude extracts via centrifugation or magnetic columns and showed enhanced stability. This study presents a modular strategy for the in vivo production of MPAs as scaffold for enzyme immobilization. The approach eliminates the need for traditional, expensive carriers and simplifies the purification process by leveraging the insoluble nature and the magnetic properties of the aggregates, opening up the potential for novel, streamlined applications in biocatalysis.

Synergistic effect between biochar and nitrate fertilizer facilitated arsenic immobilization in an anaerobic contaminated paddy soil

Nitrate nitrogen fertilizer was usually used to mitigate arsenic (As) release and mobilization in the anaerobic contaminated paddy soil. However, the effect of the interplay between nitrate fertilizer and biochar on As availability as well as the involved mechanism were poorly understood. Herein, the effects and mechanisms of biochar, nitrate fertilizer, and biochar-based nitrate fertilizer on the availability of As in the contaminated paddy soil were investigated via a microcosm incubation experiment. Results indicated that the application of biochar-based nitrate fertilizer significantly lessened the available As concentration in the contaminated paddy soil from 3.01 ± 0.03 (control group) to 2.24 ± 0.08 mg kg−1, which presented an immobilization efficiency of 26.6 % better than those of individual biochar (13.5 %) and nitrate fertilizer (17.6 %), exhibiting a synergistic effect. Moreover, the biochar-based nitrate fertilizer also facilitated the transformation of more toxic arsenite in the contaminated soil to less toxic arsenate. Further, biochar-based nitrate fertilizer increased soil redox potential (Eh), dissolved organic carbon, organic matter, and nitrate yet decreased soil pH and ammonium, which changed the microbial community in the soil, enhancing the relative abundance of Bacillus, Arthrobacter, and Paenibacillus. These functional microorganisms drove the coupled transformation between nitrate denitrification and Fe(II) or As(III) oxidation, favoring As immobilization in the anaerobic paddy soil. Additionally, the co-application of biochar offset the negative effect of single nitrate fertilizer on microbial community diversity. Overall, biochar-based nitrate fertilizer could be a promising candidate for the effective immobilization of As in the anaerobic paddy soil. The current research can provide a valuable reference to the remediation of As-contaminated paddy soil and the production of safe rice.

Mechanochemical Synthesis of High-Entropy MOF-74 with Multiple Active Sites for CO2 Adsorption and Synergistic Conversion.

Compared with monometallic metal-organic frameworks (MOFs), the synergistic effect of multiple metals significantly enhances the catalytic performance of the CO2 cycloesterification reaction, leading to improved CO2 adsorption and catalytic conversion capabilities. To investigate this concept, a high-entropy MOF-74 (HE-MOF-74) with a uniform distribution of five distinct metal ions (Zn2+, Mg2+, Ni2+, Co2+, and Cu2+) was successfully synthesized using a straightforward mechanical ball milling technique and comprehensively characterized (including structural, morphological, and physicochemical properties). The results reveal that HE-MOF-74 exhibits significantly increased specific surface area and CO2 adsorption capacity compared with those of monometallic MOF-74. The presence of multiple unsaturated metal centers as Lewis acid sites, oxygen atoms linking the metals, and ligand-based hydroxyl groups serving as base sites enable efficient immobilization of CO2 into cyclic carbonate. This study introduces a novel synthetic approach for the green and efficient production of HE-MOF-74 and proposes a new application for CO2 utilization.

Leaching Studies for the Assessment of Individual Immobilization Potential of Chrysopogon zizanioides Root Micro Powder from Lead, Cadmium and Chromium Contaminated Soil

ABSTRACT Contemporary issues of ground contamination, exacerbated by elevated heavy metal levels due to industrial activities and improper waste disposal, significantly impact soil properties, the biosphere, and result in various geotechnical problems. This study delves into the efficacy of Chrysopogon zizanioides (vetiver) root micro powder (VRMP) in immobilizing Lead (Pb), Cadmium (Cd), and Chromium (Cr) within contaminated soil to prevent groundwater pollution and bioavailability. Leaching experiments were conducted to evaluate the individual immobilization of heavy metals and determine the optimal VRMP dosage, testing four dosages (0%, 1%, 5%, and 10%) for their effectiveness in immobilizing Pb, Cd, and Cr in contaminated soil. The findings suggest that addition of VRMP led to an increase in pH, facilitating the enhanced immobilization of Pb, Cd, and Cr from the contaminated soil due to the reduction in mobility of these metals. Soil treated with 10% VRMP exhibited exceptional immobilization efficiency, achieving over 90% removal across all treatments. However, agglomeration caused the leaching process to take slightly longer. Consequently, a 5% VRMP dosage is considered optimal. In conclusion, vetiver root micro powder promotes a sustainable approach to immobilize Pb, Cd, and Cr in contaminated soils, underscoring the effectiveness of eco-friendly methodologies.

Expression and immobilization of novel N-glycan-binding protein for highly efficient purification and enrichment of N-glycans, N-glycopeptides, and N-glycoproteins.

Comprehensive and selective enrichment of N-glycans, N-glycopeptides, and N-glycoproteins prior to analysis is of great significance in N-glycomics research, reducing sample complexity, removing impurity interference, increasing sample abundance and enhancing signal intensity. However, only an Fbs1 (F-box protein that recognizes sugar chain 1) GYR variant (Fg) can enrich these N-glycomolecules solely due to its substantial binding affinity for the core pentasaccharide motif of N-glycans. Stationary phase separation is commonly used to enrich N-glycomolecules efficiently. Herein, DNA encoding the Fg was cloned into pGEX-4T-1, and the protein was expressed with a GST tag, which facilitates the convenient and efficient immobilization of recombinant GST-tagged Fg to GSH agarose resin. The yield of the GST-tagged Fg reached to 0.05 g/L after optimization of the induction condition, and the purified protein exhibited good identification ability and excellent stability for months. In particular, the immobilized GST-tagged Fg can enrich N-glycans released byPNGase Fand capture derivatized N-glycans possessing an intact terminal N-acetyl glucosamine (GlcNAc). Validation of immobilized GST-tagged Fg with standard N-glycopeptides and N-glycoproteins revealed its high loading capacity, sensitivity, and selectivity. The novel immobilized GST-tagged Fg is a convenient and efficient enrichment material specific for N-glycans, N-glycopeptides, and N-glycoproteins, suggesting excellent performance and prospects for industrial application.

Rhizosphere Effect Enhanced the Immobilization Efficiency of Mercapto-Palygorskite for Cd in Alkaline Soil: Mechanism from the Perspective of Microorganism

Rhizosphere Effect Enhanced the Immobilization Efficiency of Mercapto-Palygorskite for Cd in Alkaline Soil: Mechanism from the Perspective of Microorganism

Characteristic study of Candida rugosa lipase immobilized on lignocellulosic wastes: effect of support material.

For the first time is reported the comparison of solid biocatalysts derived from Candida rugosa lipase (CRL) immobilized on different lignocellulosic wastes (rice husk, brewer's spent grain, hemp tea waste, green tea waste, vine bark, and spent coffee grounds) focusing on the characterization of these materials and their impact on the lipase-support interaction. The wastes were subjected to meticulous characterization by ATR-FTIR, BET, and SEM analysis, besides lignin content and hydrophobicity determination. Investigating parameters influencing immobilization performance revealed the importance of morphology, textural properties, and hydrophobic interactions revealed the importance of morphology, textural properties and especially hydrophobic interactions which resulted in positive correlations between surface hydrophobicity and lipase immobilization efficiency. Hemp tea waste and spent coffee grounds demonstrated superior immobilization performances (7.20 U/g and 8.74 U/g immobilized activity, 102.3% and 33.5% efficiency, 13.4% and 15.4% recovery, respectively). Moreover, they demonstrated good temporal stability (100% and 92% residual activity after 120days, respectively) and retained 100% of their immobilized activity after five reuses in the hydrolysis of p-nitrophenyl palmitate in hexane. In addition, the study of enzymatic desorption caused by ionic strength and detergent treatments indicated mixed hydrophobic and electrostatic interactions in rice husk, vine bark, and spent coffee grounds supports, while hemp tea waste and green tea waste were dominated by hydrophobic interactions.