Investigation on solvent-free esterification of oleic acid by hemp tea waste-immobilized Candida Rugosa lipase.

This study aimed at Candida rugosa lipase immobilization on a low-cost and readily available support. Among agro-industrial crops, hemp tea waste was chosen as the carrier because it provides higher immobilization performance than hemp flower and leaf wastes. Support characterization by ATR-FTIR, SEM and elemental analysis and the optimization of the adsorption immobilization process were performed. The lipase adsorption immobilization was obtained by soaking the support with hexane under mild agitation for 2h and a successively incubating the enzyme for 1h at room temperature without removing the solvent. The esterification of oleic acid with n-decanol was tested in a solvent-free system by studying some parameters that influence the reaction, such as the substrates molar ratio, the lipase activity/oleic acid ratio, reaction temperature and the presence/absence of molecular sieves. The biocatalyst showed the ability to bring the esterification reaction to equilibrium under 60min and good reusability (maintaining 60% of its original activity after three successive esterification reactions) but low conversion (21%) at the optimized conditions (40 °C, 1:2 substrates molar ratio, 0.56 lipase/oleic acid ratio, without sieves). Comparing the results with those obtained by free lipase form at the same activity (1U) and experimental conditions, slightly higher conversion (%) appeared for the free lipase. All this highlighted that probably the source of lipase for its carbohydrate-binding pocket and lid structure affected the esterification of oleic acid but certainly, the immobilization didn't induce any lipase conformational change also allowing the reuse of the catalytic material.

Immobilized PAD4 enzyme on magnetic nanoparticles for screening natural inhibitors from traditional Chinese medicines.

Dysregulation of peptidyl arginine deiminase 4 (PAD4) is involved in a variety of diseases including rheumatoid arthritis (RA) and Alzheimer's disease (AD), and it has emerged as potential and promising therapeutic target. However, no PAD4 inhibitor is ready for clinical use. Immobilized enzyme screening technology has gained increasing attention due to its low cost, reusability, easy separation from the reaction mixture, and resistance to changes in environmental conditions. In this study, PAD4 was immobilized on the magnetic nanoparticles (MNP) to prolong its activity stability, and a simple and rapid screening strategy of traditional Chinese medicine inhibitors based on immobilized PAD4 was established. The PAD4 enzyme was immobilized on magnetic nanoparticles (MNP) via Schiff base reaction using glutaraldehyde (GA) as crosslinking agent. Compared with free PAD4, the resulting MNP@GA@PAD4 exhibited an enhanced tolerance to temperature and storage stability, and its reusability was greatly improved with 66 % of initial enzyme activity after being recycled 10 times. The inhibitory activity of the immobilized PAD4 was assessed using two known PAD4 inhibitors GSK484 and BB-Cl-amidine. The semi-maximum inhibitory concentrations (IC50) of GSK484 and BB-Cl-amidine for MNP@GA@PAD4 were 1.00 and 0.97 µM, respectively, for free PAD4 were 0.64 and 0.85 µM, respectively. Finally, the MNP@GA@PAD4 was employed to rapid screen of natural PAD4 inhibitors from forty traditional Chinese medicines (TCMs). Under the same conditions, the controlled experiment was conducted with free PAD4. The screening results of TCMs inhibitors on MNP@GA@PAD4 and free PAD4 were similar, the alcohol extracts of Cinnamomi Cortex and Caryophylli Flos had significant inhibitory effects on PAD4 enzyme activity. The IC50 values of Cinnamomi Cortex extract for MNP@GA@PAD4 and free PAD4 were determined as 27 and 48 µg/mL, respectively. The IC50 values of Caryophylli Flos extracts for MNP@GA@PAD4 and free PAD4 were determined as 48 and 32 µg/mL, respectively. For the first time, this study proposed a method to immobilize PAD4 on magnetic materials, and developed a rapid, reusable and feasible strategy to screening natural PAD4 inhibitors from TCMs.

Recent developments in the application of immobilized artificial membrane (IAM) chromatography to drug discovery.

INTRODUCTION: Immobilized artificial membrane (IAM) chromatography is widely used in many aspects of drug discovery. It employs stationary phases, which contain phospholipids combining simulation of biological membranes with rapid measurements. AREAS COVERED: Advances in IAM stationary phases, chromatographic conditions and the underlying retention mechanism are discussed. The potential of IAM chromatography to model permeability and drug-membrane interactions as well as its use to estimate pharmacokinetic properties and toxicity endpoints including ecotoxicity, is outlined. Efforts to construct models for prediction IAM retention factors are presented. EXPERT OPINION: IAM chromatography, as a border case between partitioning and binding, has broadened its application from permeability studies to encompass processes involving tissue binding. Most IAM-based permeability models are hybrid models incorporating additional molecular descriptors, while for the estimation of pharmacokinetic properties and binding to off targets, IAM retention is combined with other biomimetic properties. However, for its integration into routine drug discovery protocols, reliable IAM prediction models implemented in relevant software should be developed, to enable its use in virtual screening and the design of new molecules. Conversely, preparation of new IAM columns with different phospholipids or mixed monomers offers enhanced flexibility and the potential to tailor the conditions according to the target property.

Prevalence and Risk Factors of Pressure Ulcers in Bedbound Diabetic vs. Non-diabetic Patients in a Lower-Middle-Income Country.

Introduction Pressure ulcers, also known as bedsores, are a significant concern for bedridden individuals, presenting both physical and socioeconomic challenges. Factors such as prolonged immobility, chronic medical conditions, and poor nutrition contribute to their development. Despite extensive research in some regions, studies comparing diabetic and non-diabetic populations remain limited, particularly in low-income settings. This study aimed to investigate the risk factors and frequency of pressure ulcers among bedridden patients, addressing this gap in understanding and guiding targeted interventions. Materials and methods A cross-sectional study was conducted across four government hospitals in Peshawar, Pakistan. A total of 388 bedridden patients with pressure ulcers were included, and data were collected through a questionnaire. The questionnaire covered demographics, comorbidities, duration of bedbound status, BMI, and caregivers' awareness of pressure ulcer care. Data analysis was performed using SPSS version 22.0 (Armonk, NY: IBM Corp.), with qualitative data presented as frequencies and percentages and quantitative data as mean and standard deviation. Chi-square tests were utilized for significance, with p<0.05 considered significant. Results Of the 388 patients analyzed, 230 (59.3%) were diabetic, highlighting the prevalence of diabetes among pressure ulcer cases. The majority of diabetic patients with ulcers were over 41 years old, and 293 (75.5%) had comorbidities. Surgical intervention was the primary cause of ulcers in 213 (54.8%) cases, followed by stroke in 77 (19.8%) cases. Notably, 252 (65%) of caregivers exhibited inadequate knowledge regarding ulcer care. Stage II ulcers were prevalent in both diabetic and non-diabetic cohorts. Conclusions Pressure ulcers are poorly controlled complications observed in bedridden individuals, highlighting a critical need for comprehensive preventive measures and caregiver education to alleviate the burden of pressure ulcers, especially in diabetic patients. Factors such as prolonged immobility, surgical interventions, and insufficient caregiver knowledge contribute to the development of pressure ulcers. Understanding these complexities is essential for implementing effective care approaches and mitigating the impact of pressure ulcers.

Research trends in functionalized Fe3O4 composites based on affinity recognition systems for targeted extraction of natural products.

In recent years, target-specific affinity recognition systems based on Fe3O4-based composites have proven to be an effective method for screening natural products. Herbal medicines contain a wide range of natural products and are considered to be a major source for the development of novel drugs. However, the process of isolating and obtaining these bioactive components for the production of novel drugs is complex. Meanwhile, the complexity and diversity of herbal constituents have posed a great challenge to the screening studies of herbal active ingredients. Currently, traditional extraction and screening studies of active ingredients in herbal medicine include extraction and chromatographic separation technology development, serum medicinal chemistry, metabolomics and computerized virtual screening. In order to achieve integrated targeting of Fe3O4 for extraction and separation of natural products from herbs, various Fe3O4-based composites need to be synthesized so that the composites can be further functionalized and modified. Composites such as Fe3O4@SiO2, Fe3O4-based magnetic graphene oxide and Fe3O4-based magnetic carbon nanotubes were used to achieve targeted extraction and isolation of natural products from herbal medicines. The main extraction techniques involved based on these Fe3O4-based composites are molecularly imprinted techniques, immobilized ligand fishing techniques, and cell membrane-coated bionanotechnology methods. This article will present recent advances in the synthesis and modification of Fe3O4 composites and their applications for the extraction of natural products in conjunction with molecular imprinting, immobilization-targeted fishing, and cell-membrane-coated biomimetic techniques, as well as the future goals and challenges of functionalized modification of Fe3O4 composites for the targeted extraction of natural products, like protein overexpression modification, doping of fluorescent substances and genetic engineering development. A deeper understanding of the multi-level, multidisciplinary, and applied studies in materials science and phytochemistry will be provided by this article.

Enhanced flexibility of high-yield bamboo pulp fibers via cellulase immobilization within guar gum/polyacrylamide/polydopamine interpenetrating network hydrogels.

Softness is a crucial criterion in assessing the comfort and usability of tissue paper. Flexible fibers contribute to the softness of the tissue paper by allowing the sheets to conform to the contours of the skin without feeling rough or abrasive. This study focuses on developing innovative CGG/APAM/PDA hydrogels with interpenetrating networks consisting of cationic guar gum, anionic polyacrylamide, and polydopamine for cellulase immobilization, aimed at improving bamboo fiber flexibility. Cellulase biomolecules are efficiently immobilized on CGG/APAM/PDA hydrogels through the Schiff base reaction. Immobilized cellulases have a wider pH applicability than free cellulases, good storage stability, and can maintain high relative activity at relatively high temperatures. The treatment of bamboo fibers with immobilized cellulase results in a significant increase in flexibility, reaching 6.90 × 1014 N·m2, which is 7.18 times higher than that of untreated fibers. The immobilization of cellulases using CGG/APAM/PDA hydrogels as carriers results in a substantial enhancement of storage stability, pH applicability, and inter-fiber bonding strength, as well as the capacity to sustain high relative enzymatic activity at elevated temperatures. The immobilization of cellulase within CGG/APAM/PDA interpenetrating network hydrogels presents a viable strategy for enhancing bamboo fiber flexibility, thereby expanding the accessibility of tissue products.

Screening for promising multi-target bioactive components from Cortex Mori Radicis for the treatment of chronic cor pulmonale based on immobilized beta1-adrenergic receptor and beta2-adrenergic receptor chromatography.

Cortex Morin Radicis (CMR) is the dried root bark of Morus alba. L. It has a variety of effects such as antibacterial, anti-tumour, treatment of cardiovascular diseases or upper respiratory tract disease and so on. The pursuit for drugs selected from Cortex Mori Radicis having improved therapeutic efficacy necessitates increasing research on new assays for screening bioactive compounds with multi-targets. In this work, we applied immobilized ß1-AR and ß2-AR as the stationary phase in chromatographic column to screen bioactive compounds from Cortex Morin Radicis. Specific ligands of the two receptors (e.g. esmolol, metoprolol, atenolol, salbutamol, methoxyphenamine, tulobuterol and clorprenaline) were utilized to characterize the specificity and bioactivity of the columns. We used high performance affinity chromatography coupled with ESI-MS to screen targeted compounds of Cortex Morin Radicis. By zonal elution, we identified morin as a bioactive compound simultaneously binding to ß1-AR and ß2-AR. The compound exhibited the association constants of 3.10 × 104 and 2.60 × 104 M-1 on the ß1-AR and ß2-AR column. On these sites, the dissociation rate constants were calculated to be 0.131 and 0.097 s-1. Molecular docking indicated that the binding of morin to the two receptors occurred on Asp200, Asp121, and Val122 of ß1-AR, Asn312, Thr110, Asp113, Tyr316, Gly90, Phe193, Ile309, and Trp109 of ß2-AR. Likewise, mulberroside C was identified as the bioactive compound binding to ß2-AR. The association constants and dissociation rate constants were calculated to be 1.08 × 104 M-1 and 0.900 s-1. Molecular docking also indicated that mulberroside C could bind to ß2-AR receptor on its agonist site. Taking together, we demonstrated that the chromatographic strategy to identify bioactive natural products based on the ß1-AR and ß2-AR immobilization, has potential for screening bioactive compounds with multi-targets from complex matrices including traditional Chinese medicines.

Application of hydrophobic eutectic solvent in efficient biotransformation of total flavonoids of Herba Epimedii.

Icaritin, a hydrolysate from total flavonoids of Epimedii (TFE), which has better anti-hepatoma activity than its glycosylated form. In this work, immobilized enzymes 4LP-Tpebgl3@Na-Y and DtRha@ES-107 were used to hydrolyze TFE to prepare icaritin. Five different hydrophobic deep eutectic solvents (HDES) were prepared and the most ideal HDES was successfully selected, which was composed of dodecyl alcohol and thymol with the molar ratio of 2:1. The relative enzyme activity of 4LP-Tpebgl3@Na-Y and DtRha@ES-107 was about 102.4 % and 112.5 %, respectively. In addition, the thermal and binding stability of 4LP-Tpebgl3@Na-Y and DtRha@ES-107 in HDES was not affected negatively. In the biphasic system composed of 50 % (v/v) HDES and Na2HPO4-citric acid buffer (50 mM, pH 5.5), 4LP-Tpebgl3@Na-Y (1.0 U/mL) and TFE (1 g/L) were reacted at 80 °C for 1 h, and then reacted with DtRha@ES-107 (20 U/mL) at 80 °C for 2 h. Finally, TFE was completely converted to 301.8 mg/L icaritin (0.82 mM). After 10 cycles, 4LP-Tpebgl3@Na-Y/DtRha@ES-107 still maintained 84.1 % original activity. In this study, we developed an efficient methodology for icaritin preparation through the integration of enzymatic catalysis and adsorption separation, presenting a viable approach for large-scale, cost-effective production of icaritin.

Enhanced denitrification and p-nitrophenol removal performance via hydrophilic sponge carriers fixed with dual-bacterial: Optimization, performance, and enhancement mechanism.

Effective treatment of industrial wastewater containing complex pollutants, such as nitrate (NO3--N) and organic pollutants, remains a significant challenge to date. Here, a strain Nocardioides sp. ZS2 with denitrification and degradation of p-nitrophenol (PNP) was isolated and its culture conditions were optimized by kinetic analysis. Hydrophilic sponge carriers were prepared using polyvinyl alcohol (PVA), carboxymethyl cellulose (CMC), and chitosan (CS) to construct bioreactors. Furthermore, to further enhance the PNP degradation and denitrification performance of bioreactors, Pseudomonas stutzeri GF2 with denitrification capability was introduced. The results revealed that the removal efficiencies of PNP and NO3--N reached 97.9 % and 91.9 %, respectively, when hydraulic retention time (HRT) of 6 h, C/N of 2.0, and pH of 6.5. The bioreactor exhibited stable denitrification performance even with fluctuations in the influent PNP concentration. The potential functional prediction results revealed that the abundance of amino acids, fatty acids, and carbohydrates increased as the influent C/N decreased, reflecting a tendency of the microbial community to adjust carbon source utilization to maintain cell growth, metabolic balance, and resist adverse C/N environments. This research provides new insights into the effective removal of organic pollutants and NO3--N in wastewater treatment.

Development of automated proteomic workflows utilizing silicon-based coupling agents.

Automated methods for enzyme immobilization via 4-triethoxysilylbutyraldehyde (TESB) derived silicone-based coupling agents were developed. TESB and its oxidized derivative, 4-triethoxysilylbutanoic acid (TESBA), were determined to be the most effective. The resulting immobilized enzyme particles (IEPs) displayed robustness, rapid digestion, and immobilization efficiency of 51 ± 8%. Furthermore, we automated the IEP procedure, allowing for multiple enzymes, and/or coupling agents to be fabricated at once, in a fraction of the time via an Agilent Bravo. The automated trypsin TESB and TESBA IEPs were shown to rival a classical in-gel digestion method. Moreover, pepsin IEPs favored cleavage at leucine (>50%) over aromatic and methionine residues. The IEP method was then adapted for an in-situ immobilized enzyme microreactor (IMER) fabrication. We determined that TESBA could functionalize the silica capillary's inner wall while simultaneously acting as an enzyme coupler. The IMER digestion of bovine serum albumin (BSA), mirroring IEP digestion conditions, yielded a 33-40% primary sequence coverage per LC-MS/MS analysis in as little as 15 min. Overall, our findings underscore the potential of both IEP and IMER methods, paving the way for automated analysis and a reduction in enzyme waste through reuse, thereby contributing to a more cost-effective and timely study of the proteome. SIGNIFICANCE: This research introduces 4-triethoxysilylbutyraldehyde (TESB) and its derivatives as silicon-based enzyme coupling agents and an automated liquid handling method for bottom-up proteomics (BUP) while streamlining sample preparation for high-throughput processing. Additionally, immobilized enzyme particle (IEP) fabrication and digestion within the 96-well plate allows for flexibility in protocol where different enzyme-coupler combinations can be employed simultaneously. By enabling the digestion of entire microplates and reducing manual labor, the proposed method enhances reproducibility and offers a more efficient alternative to classical in-gel techniques. Furthermore, pepsin IEPs were noted to favor cleavage at leucine residues which represents an interesting finding when compared to the literature that warrants further study. The capability of immobilized enzyme microreactors (IMER) for rapid digestion (in as little as 15 min) demonstrated the system's efficiency and potential for rapid proteomic analysis. This advancement in BUP not only improves efficiency, but also opens avenues for a fully automated, mass spectrometry-integrated proteomics workflow, promising to expedite research and discoveries in complex biological studies.

Construction of Immobilized Laccase System Based on ZnO and Degradation of Mesotrione.

Mesotrione (MES) is a new environmental pollutant. Some reports have indicated that microbial enzymes could be utilized for MES degradation. Laccase is a green biocatalyst whose potential use in environmental pollutant detoxification has been considered limited due to its poor stability and reusability. However, these issues may be addressed using enzyme immobilization. In the present study, we sought to optimize conditions for laccase immobilization, to analyze and characterize the characteristics of the immobilized laccase, and to compare its enzymatic properties to those of free laccase. In addition, we studied the ability of laccase to degrade MES, and analyzed the metabolic pathway of MES degradation by immobilized laccase. The results demonstrated that granular zinc oxide material (G-ZnO) was successfully used as the carrier for immobilization. G-ZnO@Lac demonstrated the highest recovery of enzyme activity and exhibited significantly improved stability compared with free laccase. Storage stability was also significantly improved, with the relative enzyme activity of G-ZnO@Lac remaining at about 54% after 28 days of storage (compared with only 12% for free laccase). The optimal conditions for the degradation of MES by G-ZnO@Lac were found to be 10 mg, 6 h, 30 °C, and pH 4; under these conditions, a degradation rate of 73.25% was attained. The findings of this study provide a theoretical reference for the laccase treatment of 4-hy-droxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicide contamination.

Enzymatic synthesis and sensory evaluation of the novel kokumi compound N-butyryl phenylalanine.

In this study, food-grade glutamine transaminase (TGase) was utilized for the green-catalyzed preparation of N-butyryl amino acids. For improving the reusability of the enzyme preparation, immobilized TG enzyme (94.23% immobilization rate) was prepared. Furthermore, the yield of N-butyryl phenylalanine (BP) synthesized by TGase was obtained as 20.73% by one-factor experiment. The BP synthesis yield of immobilized TGase was 95.03% of that of TGase and remained above 60% of the initial enzyme activity after five runs. The sensory evaluation and E-tongue results showed that the addition of BP significantly increased the umami, saltiness, and richness intensities of the samples, and decreased the intensities of sourness, bitterness, and aftertaste-B. The molecular docking results indicated that hydrogen bonding dominated the binding of BP to taste receptors in the taste presentation mechanism of BP. These results confirmed the potential of BP as a flavor enhancer with promising applications in the food industry.

Purification of Recombinant N-terminal Histidine-Tagged Arabidopsis thaliana Phosphoglycolate Phosphatase 1, Glycolate Oxidase 1 and 2, and Hydroxypyruvate Reductase 1.

To measure the kinetic properties of photorespiratory enzymes, it is necessary to work with purified proteins. Protocols to purify photorespiratory enzymes from leaves of various plant species require several time-consuming steps. It is now possible to produce large quantities of recombinant proteins in bacterial cells. They can be rapidly purified as histidine-tagged recombinant proteins by immobilized metal affinity chromatography using Ni2+-NTA-agarose. This chapter describes protocols to purify several Arabidopsis thaliana His-tagged recombinant photorespiratory enzymes (phosphoglycolate phosphatase, glycolate oxidase, and hydroxypyruvate reductase) from Escherichia coli cell cultures using two bacterial strain-plasmid systems: BL21(DE3)-pET and LMG194-pBAD.

Enhancing immobilized Chlorella vulgaris growth with novel buoyant barium alginate bubble beads.

Microalgae immobilization in alginate beads shows promise for biomass production and water pollution control. However, carrier instability and mass transfer limitations are challenges. This study introduces buoyant barium alginate bubble beads (BABB), which offer exceptional stability and enhance Chlorella vulgaris growth. In just 12 days, compared to traditional calcium alginate beads, BABB achieved a 20 % biomass increase while minimizing cell leakage and simplifying harvesting. BABB optimization involved co-immobilization with BG-11 medium, enrichment of CO2 in internal bubbles, and the integration of Fe nanoparticles (FeNPs). In the open raceway pond reactor, these optimizations resulted in a 39 % increase in biomass over 7 days compared to the unoptimized setup in closed flasks. Furthermore, enhancements in pigment and organic matter production were observed, along with improved removal of ammonia nitrogen and phosphate. These results highlight the overall advantages of BABB for microalgae immobilization, offering a scientific foundation for their effective utilization.

A simple and high-performance cellulose nanocrystal based biocatalytic Pickering emulsion system for pharmaceutical molecule vitamin E nicotinate.

The use of Pickering emulsions for biocatalysis is gaining increased attention. However, the extensive application is greatly limited due to the enzyme inactivation. Herein, a biocatalytic Pickering emulsion with high-performance utilizing cellulose nanocrystals immobilized lipases (CNCs-Lps) particles as stabilizer is advanced and applied for the synthesis of Vitamin E nicotinate. CNCs-Lps display high activity and reusability due to the construction of biocatalytic microreactor in the O/W emulsion system. The yield of vitamin E nicotinate ester reached up to 83 %. More importantly, the CNCs-Lps can be reused due to the similar principles to microreactors in Pickering emulsions. Reusability test showed that the CNCs-Lps could be recovered from the emulsion system by centrifugation and the yield of vitamin E nicotinate retains 78 % of initial value after five cycles, demonstrating overwhelming advantage than the fair counterpart with free lipases.

Microorganisms immobilized hydroxyethyl cellulose/luffa composite sponge for selective adsorption and biodegradation of oils in wastewater.

The highly efficient removal of oils such as oils or dyes from wastewater has aroused wide concern and is of great significance for clean production and environmental remediation. The synthesis of a novel aerogel (designated as HEC/LS) is reported herein, achieved through a sol-gel method followed by freeze-drying utilizing loofa and hydroxyethyl cellulose as the raw materials. The new HEC/LS aerogel exhibits excellent porosity and specific surface area, with a porosity of 88.70 %, a total pore area of 0.607 m2 g-1, and a specific surface area of 230 m2 g-1. The prepared HEC/LS aerogel exhibits exceptional hydrophilicity and self-floatability, facilitating its rapid absorption of water up to 21 times its own weight within a mere 3 s. Additionally, it demonstrates good adsorption performance for methylene blue (MB), with a maximum adsorption capacity of 83.30 mg g-1. Subsequently, a new hydrophobic microorganisms-loaded composite aerogel (namely, Bn-HEC/LS) was obtained by doping microorganisms into the as-prepared HEC/LS in multiple enrichment followed by a hydrophobic and oleophilic surface modification. Based on its rich porous structure and oleophilic wettability, the as-synthesized Bn-HEC/LS exhibits excellent selective adsorption and degradation properties for the oil contamination, the diesel oil could be selectively absorbed in the Bn-HEC/LS and degraded by the loaded microorganisms. Among them, B5-HEC/LS displays the highest removal efficiency of 94.50 % within 180 h, while free microorganisms and HEC/LS aerogels show degradation efficiencies of only 21.70 % and 48.10 %, respectively. The fixation of microorganisms in the aerogel increases their number within the material and enhances the relative microorganisms removal capacity. The hydrophobic and lipophilic modifications improve the selective adsorption performance of the aerogel on diesel oil, resulting in a significantly high removal rate of Bn-HEC/LS for diesel oil. The results indicate that the immobilization of microorganisms into aerogel improves the activity of microorganisms, and the hydrophobic and oleophilic modification enhances the selective adsorption performance of aerogel to diesel oil, thus resulting in a very high removal rate of Bn-HEC/LS for diesel oil. This study is expected to provide a now possibility for the green and efficient bioremediation of oils.

Titania-zeolite composite for tetracycline photocatalytic degradation under visible light: A comparison between doping and ion exchange.

In this study, TiO2 supported over embryonic Beta zeolite (BEA) was prepared for the photocatalytic degradation of Tetracycline (TC) antibiotic under visible light. The immobilization of sol-gel TiO2 over the zeolite increased its surface area from 33 (m2/g) to 226 (m2/g) and enhanced its adsorption efficiency from 8 % to 18 %. In order to expand the photocatalytic activity of TiO2 towards the visible light region (i.e. λ > 380 nm), two different metal sensitization techniques with Iron ions from aqueous solution of FeCl3 were explored. In the ion-exchange method, the substitutional cations within the TiO2/BEA structure were exchanged with Fe3+. Whereas, in the doping technique, solgel TiO2 was doped with Fe3+ during its synthesis and before its immobilization over Zeolite. Four different samples with 20, 40, 60, and 100 % w/w of TiO2/BEA ratio were prepared. After testing the various ion-exchanged photocatalysts under blue and white lights, only Fe-60%TiO2/BEA showed better activity compared to pure TiO2 under white light at TC initial concentration, C o = 20 ppm. For the doped immobilized Titania with 60 wt% TiO2/BEA, three different doped photocatalysts were prepared with 3 %, 7 %, and 10 % per mole Fe/TiO2. All the Fe-doped TiO2/BEA photocatalysts showed better activity compared to pure TiO2 under white light. Under solar irradiations, the 3 % Fe-doped TiO2/BEA was able to degrade all TC within 120 min, while Fe-60%TiO2/BEA needed 200 min, and TiO2 needed more than 300 min. This enhanced performance was a result of both increased surface area due to immobilization over BEA as well as iron doping by Fe3+ that simultaneously increased the visible light absorption of TiO2 and minimized the charge carrier recombination effect.

Low-Temperature and High-Efficiency Solid-Phase Amplification Based on Formamide.

The thermal stability of DNA immobilized on a solid surface is one of the factors that affects the efficiency of solid-phase amplification (SP-PCR). Although variable temperature amplification ensures high specificity of the reaction by precisely controlling temperature changes, excessively high temperatures during denaturation can negatively affect DNA stability. Formamide (FA) enables DNA denaturation at lower temperatures, showing potential for SP-PCR. Research on FA's impacts on DNA microarrays is still limited, necessitating further optimization in exploring the characteristics of FA in SP-PCR according to particular application needs. We immobilized DNA on a chip using a crosslinker and generated DNA microarrays through bridge amplification based on FA denaturation on our automated reaction device. We optimized the denaturation and hybridization parameters of FA, achieving a maximum cluster density of 2.83 × 104 colonies/mm2. Compared to high-temperature denaturation, FA denaturation required a lower template concentration and milder reaction conditions and produced higher cluster density, demonstrating that FA effectively improves hybridization rates on surfaces. Regarding the immobilized DNA stability, the FA group exhibited a 45% loss of DNA, resulting in a 15% higher DNA retention rate compared to the high-temperature group, indicating that FA can better maintain DNA stability. Our study suggests that using FA improves the immobilized DNA stability and amplification efficiency in SP-PCR.

Bioremediation of Hazardous Pollutants Using Enzyme-Immobilized Reactors.

Bioremediation uses the degradation abilities of microorganisms and other organisms to remove harmful pollutants that pollute the natural environment, helping return it to a natural state that is free of harmful substances. Organism-derived enzymes can degrade and eliminate a variety of pollutants and transform them into non-toxic forms; as such, they are expected to be used in bioremediation. However, since enzymes are proteins, the low operational stability and catalytic efficiency of free enzyme-based degradation systems need improvement. Enzyme immobilization methods are often used to overcome these challenges. Several enzyme immobilization methods have been applied to improve operational stability and reduce remediation costs. Herein, we review recent advancements in immobilized enzymes for bioremediation and summarize the methods for preparing immobilized enzymes for use as catalysts and in pollutant degradation systems. Additionally, the advantages, limitations, and future perspectives of immobilized enzymes in bioremediation are discussed.

Enhanced remediation of chlorpyrifos-contaminated soil by immobilized strain Bacillus H27.

Chlorpyrifos is a pesticide widely used in agricultural production with a relatively long residual half-life in soil. Addressing the problem of residual chlorpyrifos is of universal concern. In this study, rice hull biochar was used as an immobilized carrier to prepare the immobilized strain H27 for the remediation of chlorpyrifos-contamination soil. Soil microorganisms after remediation were investigated by ecotoxicological methods. The immobilized strain H27 had the highest removal rate of chlorpyrifos when 10% bacterial solution was added to the liquid medium containing 0.075-0.109 mm diameter biochar cultured for 22 hr. This study on the removal of chlorpyrifos by immobilized strain H27 showed that the initial concentration of chlorpyrifos in solution was 25 mg/L, and the removal rate reached 97.4% after 7 days of culture. In the soil, the removal rate of the immobilized bacteria group increased throughout the experiment, which was significantly higher than that of the free bacteria and biochar treatment groups. The Biolog-ECO test, T-RFLP and RT-RCR were used to study the effects of the soil microbial community and nitrogen cycling functional genes during chlorpyrifos degradation. It was found that ICP group had the highest diversity index among the four treatment groups. The microflora of segment containing 114 bp was the dominant bacterial community, and the dominant microflora of the immobilized bacteria group was more evenly distributed. The influence of each treatment group on ammonia-oxidizing bacteria (AOB) was greater than on ammonia-oxidizing archaea (AOA). This study offers a sound scientific basis for the practical application of immobilized bacteria to reduce residual soil pesticides.