查找使用阿拉丁产品的科研文献

A novel efficient medium for chromogenic catalysis of tetramethylbenzidine with horseradish peroxidase

Meng Li,Xiang-Rong Huang,Yi Guo,Ya-Zhuo Shang,Hong-Lai Liu

The traditional surfactant sodium dodecyl sulfate (SDS) and ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate ([Emim][BF 4 ]) have been combined to create a novel efficient medium for chromogenic catalysis of 3,3′,5,5′-tetramethylbenzidine with horseradish peroxidase in presence of H 2 O 2 . The results have shown the [Emim][BF 4 ] in the mediums can promote the rate of formation of the blue chromogen, the SDS is responsible for the stabilization of the blue chromogen due to the electrostatic attraction between positively charged blue chromogen and the negatively charged surfactant. The SDS/[Emim][BF 4 ] combination not only enhance catalytic activity of HRP remarkably but also stabilize the blue chromogen formed in the HRP oxidation of the substrate TMB compared to the conventional medium. Based on the superior combination of SDS and [Emim][BF 4 ], the colorimetric assay for detecting HRP activity and H 2 O 2 concentration was established. This work demonstrates a novel efficient medium for chromogenic catalysis with potential applications in biosensors and clinical diagnosis.

A Portable Personal Glucometer-Based Immunoassay Platform for Horseradish Peroxidase-Related Biomarker Detection

Given the prevalent use of horseradish peroxidase (HRP) as an enzyme label in immunoassays, the practical importance of point-of-care testing for the portable identification of HRP-related biomarkers is evident. Accordingly, this study devised a portable immunoassay platform using a personal glucometer (PGM) to achieve rapid detection of HRP-related biomarkers. 3,3′,5,5′-tetramethylbenzidine (TMB) and dopamine (DA) were used as signal-regulating substrates, and the assay principle was based on the HRP-mediated redox reaction. In the absence of HRP, TMB and DA directly reacted with [Fe(CN)6]3−on commercial glucose test strips, producing a strong PGM response. When HRP and H2O2were present, HRP facilitated the oxidation of TMB to generate oxidized TMB, which further reacted with DA to produce oxidized products with weak reactivity toward [Fe(CN)6]3−, resulting in a decreased PGM signal. Conducting the quantitative assay involved measuring the HRP-dependent change in the PGM readout. Unlike many PGM immunoassays that rely on additional glucose-producing enzyme labels or amplified nanomaterial tags, the proposed method directly converts the signal of the conventional HRP label into a PGM-readable response. The sensing platform demonstrated a good linear response to HRP concentrations ranging from 156 to 625 ng/mL, with a low detection limit of 26 ng/mL. Interference experiments indicated good tolerance to the tested nonspecific proteins. The preliminary applicability of the platform was further evaluated by detecting cardiac troponin I (cTnI) and heparin-binding protein (HBP) in spiked serum samples, with recoveries ranging from 93.0% to 117.1% for cTnI and from 88.3% to 112.5% for HBP. In summary, the proposed platform provides a portable digital readout for HRP-linked immunoassays and reduces the dependence on dedicated readout instruments.

Electrochemical Biosensor Based on Horseradish Peroxidase and Black Phosphorene Quantum Dot Modified Electrode

Xiaoqing Li,Fan Shi,Lisi Wang,Siyue Zhang,Lijun Yan,Xiaoping Zhang,Wei Sun

Black phosphorene quantum dots (BPQDs) were prepared by ultrasonic-assisted liquid-phase exfoliation and centrifugation with morphologies proved by TEM results. Furthermore, an electrochemical enzyme sensor was prepared by co-modification of BPQDs with horseradish peroxidase (HRP) on the surface of a carbon ionic liquid electrode (CILE) for the first time. The direct electrochemical behavior of HRP was studied with a pair of well-shaped voltammetric peaks that appeared, indicating that the existence of BPQDs was beneficial to accelerate the electron transfer rate between HRP and the electrode surface. This was due to the excellent properties of BPQDs, such as small particle size, high interfacial reaction activity, fast conductivity, and good biocompatibility. The presence of BPQDs on the electrode surface provided a fast channel for direct electron transfer of HRP. Therefore, the constructed electrochemical HRP biosensor was firstly used to investigate the electrocatalytic behavior of trichloroacetic acid (TCA) and potassium bromate (KBrO3), and the wide linear detection ranges of TCA and KBrO3were 4.0–600.0 mmol/L and 2.0–57.0 mmol/L, respectively. The modified electrode was applied to the actual samples detection with satisfactory results.Keywords:black phosphorene quantum dots;horseradish peroxidase;direct electrochemistry;electrochemical detection

Changes in activity, structure and morphology of horseradish peroxidase induced by cold plasma

Yong-Xu Han,Jun-Hu Cheng,Da-Wen Sun

Cold plasma is an emerging technology increasingly applied in the agri-food industry. For fruit and vegetables, enzymatic browning is a common phenomenon, causing quality deterioration. The objective of this study was to illustrate the effect of microscale atmospheric-pressure plasma jet (µAPPJ) plasma on the horseradish peroxidase (HRP). Results showed that after plasma treatment for 10 min, the residual activity of HRP was decreased to around 17%, and modification of secondary and tertiary structures were confirmed. The atomic force microscope (AFM) analysis revealed that the aggregation of enzyme protein was enhanced with prolonging treatment time. It was concluded that the activity of HRP could be reduced with destruction of structures and deformation of microstructure induced by µAPPJ plasma. The current study attempted to provide new idea for inhibiting browning enzymes of fruit and vegetables with plasma technology through deeper understanding of the interaction mechanism of plasma active species with enzymes.

Preparation of photo-crosslinkable cinnamate modified hyaluronic acid for immobilization of horseradish peroxidase

Raedah A.S. Alatawi,M. Monier,Nadia H. Elsayed

In this work, a photo-responsive hydrogel membrane based on cinnamate-modified hyaluronic acid (HA-CM) was developed and safely cross-linked under UV light curing. The obtained material was effectively utilized for immobilization of horseradish peroxidase (HRP) enzyme via encapsulation and entrapment strategy with efficiency above 95%. The prepared HA-CM biopolymer was investigated before the UV curing using instrumental and spectral techniques including Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR). During the UV irradiation, the progress of the cross-linking reaction was monitored by the UV–vis light spectroscopy. In addition, when the photo-induced cross-linking had accomplished, the morphological appearance of the hydrogel membrane was recorded using a scanning electron microscope (SEM). The HRP immobilized in HA-CM membranes displayed remarkable stability against the environmental pH changes especially under alkaline media and shift the optimum pH to 8 compared to the free HRP, which exhibited the highest activity at pH 7. Also, the entrapped enzyme was able to preserve above 85% of its catalytic activity at higher temperature values where the free enzyme had deactivated by approximately 50%. Moreover, HA-CM-HRP maintained 87% of its activity after 10 sequential reuse cycles, which indicate the economic value of the employed immobilization strategy.

Synthesis and characterization of starch-poly(methyl acrylate) graft copolymers using horseradish peroxidase

Su Wang,Qiang Wang,Xuerong Fan,Jin Xu,Ying Zhang,Jiugang Yuan,Heling Jin,Artur Cavaco-Paulo

Horseradish peroxidase (HRP)-mediated graft polymerization in the presence of hydrogen peroxide (H 2 O 2 ) and acetylacetone (Acac) has been successfully applied to the synthesis of starch-poly(methyl acrylate) (PMA). The graft copolymer was characterized by Fourier transform infrared (FT-IR), elemental analysis, nuclear magnetic resonance ( 1 H NMR and 13 C NMR), and differential scanning calorimetry (DSC). FT-IR, elemental analysis and NMR confirmed that methyl acrylate (MA) was grafted onto starch successfully. DSC results showed the graft reaction had changed the crystalline regions of the gelatinized starch. The effects of pH, MA content, HRP dosage, incubation temperature and time on grafting percentage (GP) and grafting efficiency (GE) were also investigated. The GP and GE under optimal conditions reached 30.21% and 45.13%, respectively.

Immobilization of Horseradish Peroxidase and Myoglobin Using Sodium Alginate for Treating Organic Pollutants

Xinyu Wang,Hossein Ghanizadeh,Shoaib Khan,Xiaodan Wu,Haowei Li,Samreen Sadiq,Jiayin Liu,Huimin Liu,Qunfeng Yue

Removing organic pollutants from wastewater is crucial to prevent environmental contamination and protect human health. Immobilized enzymes are increasingly being explored for wastewater treatment due to their specific catalytic activities, reusability, and stability under various environmental conditions. Peroxidases, such as horseradish peroxidase (HRP) and myoglobin (Mb), are promising candidates for immobilized enzymes utilized in wastewater treatment due to their ability to facilitate the oxidation process of a wide range of organic molecules. However, the properties of the carrier and support materials greatly influence the stability and activity of immobilized HRP and Mb. In this research, we developed immobilized HRP and Mb using support material composed of sodium alginate and CaCl2as carriers and glutaraldehyde as a crosslinking agent. Following this, the efficacy of immobilized HRP and Mb in removing aniline, phenol, and p-nitrophenol was assessed. Both immobilized enzymes removed all three organic pollutants from an aqueous solution, but Mb was more effective than HRP. After being immobilized, both enzymes became more resilient to changes in temperature and pH. Both immobilized enzymes retained their ability to eliminate organic pollutants through eight treatment cycles. Our study uncovered novel immobilized enzyme microspheres and demonstrated their successful application in wastewater treatment, paving the way for future research.

Sensitive sulfide ion detection by optofluidic catalytic laser using horseradish peroxidase (HRP) enzyme

Chaoyang Gong,Yuan Gong,Maung Kyaw Khaing Oo,Yu Wu,Yunjiang Rao,Xiaotian Tan,Xudong Fan

We report an optofluidic catalytic laser for sensitive sulfide ion detection. In the catalytic reaction , horseradish peroxidase (HRP) enzyme is used for catalyzing the non-fluorescent substrate, 10-Acetyl-3,7-dihydroxyphenox-azine (ADHP), to produce highly fluorescent resorufin , which was used as gain medium for lasing. Using sulfide ions as inhibitors, the catalytic reaction slows down, resulting in a delay in the lasing onset time, which is used as the sensing signal. The sensing mechanism of the catalytic laser is theoretically analyzed and the performance is experimentally characterized. Sulfide anion is chosen as a model ion because of its broad adverse impacts on both environment and human health. Due to the optical feedback provided by the laser, the small difference in the sulfide ion concentration can be amplified. Consequently, a detection limit of 10   nM is achieved with a dynamic range as large as three orders of magnitude, representing significant improvement over the traditional fluorescence and colorimetric methods. This work will open a door to a new catalytic-laser-based chemical sensing platform for detecting a wide range of species that could inhibit the catalytic reaction.

Chitosan as an eco-friendly template for enzymatic textile dyeing with horseradish peroxidase

Na-won Baek

Enzyme-based dyeing processes have demonstrated the efficacy of synthetic templates; however, research on natural templates remains limited. This study explores the feasibility of enzyme dyeing processes for cellulose and protein fibers and verifies the potential of chitosan (CS) as a template to replace chemical templates. The dyeing mechanism of textiles was thoroughly analyzed using liquid chromatography-tandem mass spectrometry. The dyeability and functionality of dyed textiles were examined through Fourier-transform infrared spectroscopy, thermogravimetric analysis, and scanning electron microscopy. The color and colorfastness of textiles were evaluated using color depth analysis and various fastness tests. The results demonstrated that applying CS templates in the dyeing process increased K/S values and improved colorfastness for both silk and cotton fabrics. The K/S value of the dyed silk fabric increased from 5.6 to 7.8 following the application of the chitosan template, with color fastness improving by approximately one grade. For the dyed cotton fabric, the K/S value rose from 2.2 to 3.1, and color fastness improved by one or two grades on average.

Catalytic polymerization of bisphenol A using a horseradish peroxidase immobilized microporous membrane reactor

Li Haitao,Guo Linfeng,Li Yingying,Chen Min,Bai Chunlu,Song Aolei,Cheng Linxiu,Chen Xueli,Chen Yonglin

Bisphenol A (BPA) is one of the most widely used chemical products, which is discharged into rivers and oceans, posing great hazards to organisms such as reproductive toxicity, hormone imbalance and cardiopathy induction. With the expansion harm of BPA, people have paid more attention to the environmental effects. In this paper, the degradation of BPA from the synthetic wastewater using the immobilization of horseradish peroxidase membrane reactor (HPR) was investigated. The immobilized HRP microporous membrane was prepared by the porous calcium alginate method. In addition, the reuse of the immobilized HPR membrane and the measurement of membrane flux showed that the membrane has good activity and stability. Finally, the experimental parameters including reaction time, pH, the concentration of BPA and the dosage of H 2 O 2 were optimized to remove the BPA, and about 78% degradation efficiency of BPA was achieved at the optimal condition as follows: H 2 O 2 to BPA molar ratio of 1.50 with an initial BPA concentration of 0.1 mol/L, the HPR dosage of 3.84 u/mL, the initial solution pH of 7.0, a temperature of 20 °C and a contact time of 10 min. Horseradish peroxidase (HRP) can be effectively immobilized on the membrane. The immobilized HRP can maintain high activity and be circulation utilization. The optimum conditions for the mole rate of H 2 O 2 /BPA dosage and the pH value are 1.5 and 7, respectively.

UV-Vis detection of hydrogen peroxide using horseradish peroxidase/copper phosphate hybrid nanoflowers

Chuankai Yang,Miaorong Zhang,Wei Wang,Yao Wang,Jianguo Tang

A colorimetric platform based on enzyme-inorganic hybrid nanoflowers was used for ultraviolet-visible detection of hydrogen peroxide (H 2 O 2 ). The hybrid nanoflowers were prepared via incubation at room temperature using horseradish peroxidase (HRP) and copper phosphate (Cu 3 (PO 4 ) 2 ) as the organic and inorganic components, respectively. HRP/Cu 3 (PO 4 ) 2 hybrid nanoflowers (HRP-HNFs) showed linear H 2 O 2 detection range from 100 nM to 100 μM, moreover, HRP-HNFs exhibited good selectivity for H 2 O 2 detection due to the specificity of HRP. In addition, HRP-HNFs have good reusability, excellent color stability and long-term storage stability, which indicate HRP-HNFs are promising for catalysis and biomedical applications.

Comparison of soybean peroxidase with horseradish peroxidase and alkaline phosphatase used in immunoassays

Huijuan Yang,Candace S. Bever,Huiyan Zhang,Ghulam Mujtaba Mari,Hongfang Li,Xiya Zhang,Liuchuan Guo,Ziwen Wang,Pengjie Luo,Zhanhui Wang

Enzyme labeling of an antigen or an antibody helps to visualize and amplify the signal and is an important reagent used in immunoassays for the detection of a target of interest. In this research, soybean peroxidase (SBP), a less commonly used enzyme reporter, was compared in immunoassays with the two most commonly used reagents, horseradish peroxidase (HRP) and alkaline phosphatase (ALP). The enzyme-antibody conjugates were evaluated by their performance in an indirect competitive enzyme-linked immunosorbent assay (icELISA) and in an indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) for ractopamine (RAC). The results revealed that the more affordable SBP offers a long-lasting chemiluminescent signal, which outperformed ALP and HRP. SBP-antibody conjugate (SBP-Ab) based immunoassays produced lower limits of detection (LODs) and better accuracy in the detection of RAC in animal urine samples. Additionally, SBP-Ab has advantages in being more resistant to heat, acid and organic solvents . These results suggest that SBP could be a potentially excellent alternative to HRP and ALP for the development of immunoassay in food safety field.