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1.
Xiaohua Wang  Lina Zhang   《Carbohydrate research》2009,344(16):2209-2216
Five fractions of lentinan, a β-(1→3)-d-glucan bearing β-(1→6)-d-glucopyranosyl branches, were treated with chlorosulfonic acid for 90 min at 60 °C in pyridine medium to synthesize water-soluble sulfated derivatives having the substitution degree of 1.44–1.76. The 13C NMR spectra of the sulfated β-glucans indicated that the C-6 position was preferentially substituted by the sulfate groups. The values of the weight-average molecular weight (Mw), radius of gyration (), and intrinsic viscosity ([η]) of the sulfated lentinan fractions were determined by size-exclusion chromatography with multi-angle laser light scattering (SEC–MALLS) and viscometry in 0.15 M aq NaCl at 25 °C, respectively. The dependence of [η] on Mw for the sulfated lentinan was found to be [η] = 8.93 × 10−3 (mL/g) in 0.15 M aq NaCl (for Mw ranging from 14.6 × 104 to 50.4 × 104). On the basis of the Yamakawa–Fujii–Yoshizaki (YFY) theory, the conformational parameters of the sulfated lentinan were calculated as 950 nm−1 for the molar mass per unit contour length (ML), 4.8 nm for the persistence length (q), and 13.9 for the characteristic ratio (C), indicating relatively extended single flexible chains in solution. The sulfated glucan fractions exhibited in vitro antiproliferative activities against sarcoma 180 (S-180) cells, and their inhibition ratios were lower than that of the triple-helix lentinan, but higher than that for the one with single random-coil lentinan chains.  相似文献   

2.
Photosynthesis, water relations, chlorophyll fluorescence, and leaf reflectance were used to evaluate stress due to freshwater and saltwater flooding in the evergreen coastal shrub, Myrica cerifera, under controlled conditions. M. cerifera forms large monospecific thickets that facilitate scaling up from leaf-level measurements to the landscape. Based on physiological responses, stress began by day 3 in flooded plants treated with 5, 10, and 15 g L−1 salinity, as seen by significant decreases in stomatal conductance and net photosynthesis relative to control plants. Decreases in physiological measurements occurred by day 9 in freshwater flooded plants. Visible signs of stress occurred by day 5 for plants treated with 15 g L−1, day 8 for flooded plants exposed to 10 g L−1, and day 10 for those treated with 5 g L−1 salinity. Significant differences in light-adapted fluorescence yield () were observed by day 3 in plants flooded with 5, 10, and 15 g L−1 salinity and day 6 in freshwater flooded plants. Non-photochemical quenching (ΦNPQ) increased with decreasing . In comparison, statistical differences in dark-adapted fluorescence yield (Fv/Fm) were observed by day 12 in plants flooded with 5, 10, and 15 g L−1 salinity, well after visible signs of stress were apparent. Fluorescence parameters were successful at detecting and distinguishing both freshwater and saltwater flooding stress. A positive, linear correlation (r2 = 0.80) was observed between and the physiological reflectance index (PRI). Xanthophyll-cycle dependent energy dissipation appears to be the underlying mechanism in protecting photosystem II from excess energy in saltwater flooded plants. was useful in detecting stress-induced changes in the photosystem before any visible signs of damage were evident at the leaf-level. This parameter may be linked to hyperspectral reflectance data for rapid detection of stress at the canopy-level.  相似文献   

3.
A biosensor based on the ionic liquid 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (BMI·Tf2N) and a novel source of peroxidase (tissue from the pine nuts of Araucaria angustifolia) was constructed. This enzyme was immobilized on chitosan crosslinked with citrate and the biosensor used for the determination of rosmarinic acid by square-wave voltammetry. The peroxidase in the presence of hydrogen peroxide catalyzes the oxidation of rosmarinic acid to quinone and the electrochemical reduction of the product was obtained at a potential of +0.15 V vs. Ag/AgCl. Different analytical parameters influencing the biosensor response, that is, peroxidase units, pH, hydrogen peroxide concentration and parameters for the square-wave voltammetry (frequency, pulse amplitude and scan increment), were investigated. The best performance was observed for the biosensor under the following conditions: 1000 units mL−1 peroxidase, pH 7.0 and 8.3 × 10−4 mol L−1 hydrogen peroxide with a frequency of 30 Hz, pulse amplitude of 100 mV and scan increment of 5.0 mV. The biosensor gave a linear response to rosmarinic acid over the concentration range of 9.07 × 10−7 to 4.46 × 10−6 mol L−1 with a detection limit of 7.25 × 10−8 mol L−1. The recovery of rosmarinic acid in plant extracts ranged from 97.0% to 109.6% and the determination of this substance in these samples using the biosensor compared favorably with that using the capillary electrophoresis method.  相似文献   

4.
A new acridone derivative 2-nitroacridone (NAD) was synthesized in this paper, and it was found that NAD had excellent electrochemical activity on the glassy carbon electrode (GCE) with a couple reversible redox peaks at 0.051 V and 0.103 V, respectively. Voltammetry was used to investigate the electrochemical behavior of NAD and the interaction between NAD and salmon sperm DNA. In pH 4.0 phosphate buffer solution, the binding ratio between NAD and salmon sperm DNA was calculated to be 2:1 and the binding constant was 3.19 × 105 L/mol. A Chronic Myelogenous Leukemia (CML, Type b3a2) DNA biosensor was developed by immobilizing covalently single-stranded CML DNA fragments to a modified GCE. The surface hybridization of the immobilized single-stranded CML DNA fragment with its complementary DNA fragment was evidenced by electrochemical methods using NAD as a novel electrochemical indicator, with a detection limit of 6.7 × 10−9 M and a linear response range of 1.8 × 10−8 M to 9.1 × 10−8 M for CML DNA. Selective determination of complementary ssDNA was achieved using differential pulse voltammetry (DPV).  相似文献   

5.
A sensitive, selective, and rapid enzymatic method is proposed for the quantification of hydrogen peroxide (H2O2) using 3-methyl-2-benzothiazolinonehydrazone hydrochloride (MBTH) and 10,11-dihydro-5H-benz(b,f)azepine (DBZ) as chromogenic cosubstrates catalyzed by horseradish peroxidase (HRP) enzyme. MBTH traps free radical released during oxidation of H2O2 by HRP and gets oxidized to electrophilic cation, which couples with DBZ to give an intense blue-colored product with maximum absorbance at 620 nm. The linear response for H2O2 is found between 5 × 10−6 and 45 × 10−6 mol L−1 at pH 4.0 and a temperature of 25 °C. Catalytic efficiency and catalytic power of the commercial peroxidase were found to be 0.415 × 106 M−1 min−1 and 9.81 × 10−4 min−1, respectively. The catalytic constant (kcat) and specificity constant (kcat/Km) at saturated concentration of the cosubstrates were 163.2 min−1 and 4.156 × 106 L mol−1 min−1, respectively. This method can be incorporated into biochemical analysis where H2O2 undergoes catalytic oxidation by oxidase. Its applicability in the biological samples was tested for glucose quantification in human serum.  相似文献   

6.
A water-insoluble (1 → 3)-β-d-glucan (PCSG) isolated from the fresh sclerotium of Poria cocos was carboxymethylated to afford a water-soluble derivative coded as C-PCSG. The carboxymethylated (1 → 3)-β-d-glucan was fractionated to obtain eight fractions according to the nonsolvent addition method. The weight-average molecular mass (Mw), radius of gyration and intrinsic viscosity ([η]) of the fractions were determined by size-exclusion chromatography combined with laser light scattering (SEC-LLS) and viscometry in 0.2 M NaCl aqueous solution at 25 °C. The dependences of [η] and on Mw for C-PCSG were found to be , and (nm), respectively. Analysis of Mw and [η] in terms of the known theories for wormlike chain model yielded 633 nm−1 for molar mass per unit contour length (ML), 5.5 nm for persistence length (q), and 20.2 for characteristic ratio (C). These results indicated that C-PCSG exists as a relatively extended flexible chain in 0.2 M NaCl aqueous solution. Therefore, the introduction of the carboxymethyl groups into the β-glucan improved significantly the water solubility and enhanced the stiffness of the chains.  相似文献   

7.
In this work, colloidal laponite nanoparticles were further expanded into the design of the third-generation biosensor. Direct electrochemistry of the complex molybdoenzyme xanthine oxidase (XnOx) immobilized on glassy carbon electrode (GCE) by laponite nanoparticles was investigated for the first time. XnOx/laponite thin film modified electrode showed only one pair of well defined and reversible cyclic voltammetric peaks attributed to XnOx–FAD cofactor at about −0.370 V vs. SCE (pH 5). The formal potential of XnOx–FAD/FADH2 couple varied linearly with the increase of pH in the range of 4.0–8.0 with a slope of −54.3 mV pH−1, which indicated that two-proton transfer was accompanied with two-electron transfer in the electrochemical reaction. More interestingly, the immobilized XnOx retained its biological activity well and displayed an excellent electrocatalytic performance to both the oxidation of xanthine and the reduction of nitrate. The electrocatalytic response showed a linear dependence on the xanthine concentration ranging from 3.9 × 10−8 to 2.1 × 10−5 M with a detection limit of 1.0 × 10−8 M based on S/N = 3.  相似文献   

8.
Highly ordered Ni nanowire arrays (NiNWAs) were synthesized for the first time using a template-directed electropolymerization strategy with a nanopore polycarbonate (PC) membrane template, and their morphological characterization were examined by scanning electron microscopy (SEM) and transmission electron microscope (TEM). A NiNWAs based electrode shows very high electrochemical activity for electrocatalytic oxidation of glucose in alkaline medium, which has been utilized as the basis of the fabrication of a nonenzymatic biosensor for electrochemical detection of glucose. The biosensor can be applied to the quantification of glucose with a linear range covering from 5.0 × 10−7 to 7.0 × 10−3 M, a high sensitivity of 1043 μA mM−1 cm−2, and a low detection limit of 1 × 10−7 M. The experiment results also showed that the sensor exhibits good reproducibility and long-term stability, as well as high selectivity with no interference from other oxidable species.  相似文献   

9.
An attractive biocomposite based on polycrystalline bismuth oxide (BiOx) film and polyphenol oxidase (PPO) was proposed for the construction of a mediator-free amperometric biosensor for phenolic compounds in environmental water samples. The phenolic biosensor could be easily achieved by casting the biocomposite on the surface of glassy carbon electrode (GCE) via the cross-linking step by glutaraldehyde. The laboratory-prepared bismuth oxide semiconductor was polymorphism. Its hydrophilicity provided a favorable microenvironment for retaining the biological activity of the immobilized protein. The parameters of the fabrication process and the various experimental variables for the enzyme electrode were optimized. The proposed PPO/BiOx biosensor provided a linear response to catechol over a concentration range of 4 × 10−9 M to 1.5 × 10−5 M with a dramatically developed sensitivity of 11.3 A M−1 cm−2 and a detection limit of 1 × 10−9 M based on S/N = 3. In addition, the PPO/BiOx biocomposite was characterized by scanning electron microscope (SEM), Fourier transform infrared spectra (FTIR) and rotating disk electrode voltammetry.  相似文献   

10.
Successful organ cryopreservation will significantly benefit human health and biomedical research. One of the major challenges to this accomplishment is the need for optimization of cryoprotectant agent (CPA) perfusion procedures that involve highly complicated mass transfer processes in organs. The diffusivity of CPA is of critical importance for designing perfusion procedures to minimize the associated toxicity and osmotic damage. However, to date there have been no attempts to measure the CPA diffusivity in organs. In this study, we established a simple CPA diffusion model for relatively small organs, e.g., mouse ovaries, defined the apparent diffusivity () of CPA for these organs, and established a practical approach to measure the value of through magnetic resonant imaging (MRI). Using rapid MRI techniques and water saturation analyses, the distribution of ethylene glycol (EG) concentration in the centric cross-section of mouse ovaries was measured at a series of time points during perfusion, and these data were fit to the integral form of the mass transfer equation in the established model. These fits resulted in a value of for EG in mouse ovaries of 6.1 ± 1.4 × 10−7 cm2/s (mean ± SD). Based on these results, we proposed a modified perfusion procedure that may improve the survival of small organs or thin tissues during equilibrium cooling processes and assessed its efficiency through theoretical analyses.  相似文献   

11.
We present the design, fabrication and optical investigation of photonic crystal (PhC) nanocavity drop filters for use as optical biosensors. The resonant cavity mode wavelength and Q-factor are studied as a function of the ambient refractive index and as a function of adsorbed proteins (bovine serum albumin) on the sensor surface. Experiments were performed by evanescent excitation of the cavity mode via a PhC waveguide. This in turn is coupled to a ridge waveguide that allows the introduction of a fluid flow cell on a chip. A response of ∂λ/∂c=(4.54±0.66)×105 nm/M is measured leading to a measured detection limit as good as  fg or  pg/mm2in the sensitive area.  相似文献   

12.
Carbonaceous adsorbents based on sawdust pellets from coniferous tree species were obtained by carbonisation at different temperatures and different periods of time. The effect of NO2 adsorption in dry and wet condition on the sorption ability of the chars obtained was tested. The results have shown that NO2 sorption properties of chars depend on the conditions of pyrolysis and the conditions of adsorption. The best NO2 sorption capacity of 18.3 and 43.1 mg/g in dry and wet conditions, respectively, was noted for the char sample pyrolysed at 800 °C for 60 min. The FTIR spectra of the exhausted samples reveal a great increase in the intensity of the band at 1380 cm−1 assigned to the vibrations of –NO2, –ONO2 or , while in the DTG curves a new peak appears in the range 200–400 °C assigned to the release of nitrogen compounds of low stability in high temperatures.  相似文献   

13.
Novel Pt nanoclusters embedded polypyrrole nanowires (PPy-Pt) composite was electrosynthesized on a glassy carbon electrode, denoted as PPy-Pt/GCE. A glucose biosensor was further fabricated based on immobilization of glucose oxidase (GOD) in an electropolymerized non-conducting poly(o-aminophenol) (POAP) film that was deposited on the PPy-Pt/GCE. The morphologies of the PPy nanowires and PPy-Pt nanocomposite were characterized by field emission scanning electron microscope (FE-SEM). Effect of experimental conditions involving the cycle numbers for POAP deposition and Pt nanoclusters deposition, applied potential used in glucose determination, temperature and pH value of the detection solution were investigated for optimization. The biosensor exhibited an excellent current response to glucose over a wide linear range from 1.5 × 10−6 to 1.3 × 10−2 M (r = 0.9982) with a detection limit of 4.5 × 10−7 M (s/n = 3). Based on the combination of permselectivity of the POAP and the PPy films, the sensor had good anti-interference ability to ascorbic acid (AA), uric acid (UA) and acetaminophen. The apparent Michaelis–Menten constant (Km) and the maximum current density (Im) were estimated to be 23.9 mM and 378 μA/cm2, respectively. In addition, the biosensor had also good sensitivity, stability and reproducibility.  相似文献   

14.
Accurate characterization of the biochemical pathways of nitric oxide (NO) is essential for investigations in the field of NO research. To analyze the different reaction pathways of enzymatic and non-enzymatic NO formation, determination of the source of NO is crucial. Measuring NO-related products in biological samples distinguishing between 14NO and 15NO offers the opportunity to specifically analyze NO signaling in blood and tissue. The aim of this study was to establish a highly sensitive technique for the specific measurement of NO in an isotopologue-selective manner in biological samples.With the cavity leak-out spectroscopy setup (CALOS) a differentiation between 14NO and 15NO is feasible. We describe here the employment of this method for measurements in biological samples. Certified gas mixtures of 14NO/N2 and 15NO/N2 were used to calibrate the system. and of aqueous and biological samples were reduced in a triiodide solution, and the NO released was detected via CALOS. Gas-phase chemiluminescence detection (CLD) was used for evaluation.The correlation received for both methods for the detection of NO in the gas phase was r = 0.999, p < 0.0001. Results obtained using aqueous and biological samples verified that CALOS enables NO measurements with high accuracy (detection limit for 0.3 pmol and 0.5 pmol; correlation 14NO: p < 0.0001, r = 0.975, 15NO: p < 0.0001, r = 0.969).The CALOS assay represents an extension of NO measurements in biological samples, allowing specific investigations of enzymatic and non-enzymatic NO formation and metabolism in a variety of samples.  相似文献   

15.
An ethanol biosensor, based on the electrogenerated chemiluminescence of Ru(bpy)32+-doped silica nanoparticles (RuSiNPs), was investigated in this study. The biosensor was a modified glassy carbon electrode, where alcohol dehydrogenase was crosslinked to RuSiNPs, and then immobilized on the electrode surface using chitosan. The results indicated that the biosensor exhibited excellent performance during ethanol determination with a wide linear range (10−7 to 10−2 M), low detection limit (5.0 × 10−8 M) and good stability.  相似文献   

16.
The direct immobilization of glucose oxidase (GOD) on TiO2/SiO2 nanocomposite and its application as glucose biosensor were investigated. The room-temperature phosphorescence of TiO2/SiO2 nanocomposite can be quenched by hydrogen peroxide (H2O2). The detection of glucose may be accomplished by monitoring the formation of hydrogen peroxide which generated in the oxidation process of glucose with the catalysis of GOD. To our surprise, by using a 96-hole polyporous plate accessory of fluorescence spectrophotometer, the biosensor exhibits excellent linear response to glucose concentrations ranging from 1.0 × 10−9 to 1.0 × 10−2 M with a detection limit of 1.2 × 10−10 M. The TiO2/SiO2 nanocomposite can be used as both supporting material and signal transducer. The phosphorescence intensity and color of the biosensor change obviously and even could be observed with naked eyes by continuous addition of glucose. Based on the room-temperature phosphorescence of TiO2/SiO2 nanocomposite, a new method of solid substrate-room-temperature phosphorimetry (SS-RTP) for glucose determination is proposed. A glucose biosensor was fabricated with wide determination concentration range, low detection limit, high sensitivity, and fast response time. And the biosensor has been successfully applied to the determination of glucose in human blood serum. The coacervation of GOD enzyme and its interaction with TiO2/SiO2 nanocomposite enlarge the surface area and enhance the chemical stability of GOD. The nice biocompatibility, large surface area, good chemical stability and nontoxicity of the TiO2/SiO2 nanocomposite have made this material suitable for functioning as biosensor.  相似文献   

17.
The kinetics of the esterification of oleic acid with 1-butanol catalyzed by free Rhizomucor miehei lipase in a biphasic system was studied in a batch reactor. The reaction appeared to proceed via a Ping Pong bi–bi mechanism with 1-butanol inhibition. The kinetic constants of the model were determined from experiments at 30 °C with initial concentrations of oleic acid and 1-butanol in the organic phase and 0.05–0.2 g L−1 enzyme in the aqueous phase. The model was used to simulate the batch concentration profiles of the product as well as the initial reaction rates. Agreement of the model with both the batch concentration profiles (average error of 7.2%) and the initial reaction rate per experiment (average error of 16.0%) was good.  相似文献   

18.
A novel graphitized ordered macroporous carbon (GMC, pore size 380 nm) with hierarchical mesopores (2–30 nm) and high graphitization degree was prepared by nickel-catalyzed graphitization of polystyrene arrays. The obtained GMC possessed high specific surface area, large pore volume, and good electrical conductivity, which was explored for the enzyme entrapment and biosensor fabrication by a facile method. With advantages of novel nanostructure and good electrical conductivity, direct electrochemistry of hemoglobin (a model protein) was observed on the GMC-based biocomposite with a formal potential of −0.36 V (vs. Ag/AgCl) and an apparent heterogeneous electron transfer rate constant (ks) of 1.2 s−1 in pH 7.0 buffer. Comparative studies revealed that GMC offered significant advantages over carbon nanotubes (CNTs) in facilitating direct electron transfer of entrapped Hb. The fabricated biosensor exhibited good sensitivity (101.6 mA cm−2 M−1) and reproducibility, wide linear range (1–267 μM), low detection limit (0.1 μM), and good long-term stability for H2O2 detection. GMC proved to be a promising matrix for enzyme entrapment and biosensor fabrication, and may find wide potential applications in biomedical detection and environmental analyses.  相似文献   

19.
Electrochemical oxidation of serotonin (SN) onto zinc oxide (ZnO)-coated glassy carbon electrode (GCE) results in the generation of redox mediators (RMs) that are strongly adsorbed on electrode surface. The electrochemical properties of zinc oxide-electrogenerated redox mediator (ZnO/RM) (inorganic/organic) hybrid film-coated electrode has been studied using cyclic voltammetry (CV). The scanning electron microscope (SEM), atomic force microscope (AFM), and electrochemical techniques proved the immobilization of ZnO/RM core/shell microparticles on the electrode surface. The GCE modified with ZnO/RM hybrid film showed two reversible redox peaks in acidic solution, and the redox peaks were found to be pH dependent with slopes of −62 and −60 mV/pH, which are very close to the Nernst behavior. The GCE/ZnO/RM-modified electrode exhibited excellent electrocatalytic activity toward the oxidations of ascorbic acid (AA), dopamine (DA), and uric acid (UA) in 0.1 M phosphate buffer solution (PBS, pH 7.0). Indeed, ZnO/RM-coated GCE separated the anodic oxidation waves of DA, AA, and UA with well-defined peak separations in their mixture solution. Consequently, the GCE/ZnO/RMs were used for simultaneous detection of DA, AA, and UA in their mixture solution. Using CV, calibration curves for DA, AA, and UA were obtained over the range of 6.0 × 10−6 to 9.6 × 10−4 M, 1.5 × 10−5 to 2.4 × 10−4 M, and 5.0 × 10−5 to 8 × 10−4 M with correlation coefficients of 0.992, 0.991, and 0.989, respectively. Moreover, ZnO/RM-modified GCE had good stability and antifouling properties.  相似文献   

20.
Laccase-catalyzed oxidation of phenolic compounds in organic media   总被引:1,自引:0,他引:1  
Rhus vernificera laccase-catalyzed oxidation of phenolic compounds, i.e., (+)-catechin, (−)-epicatechin and catechol, was carried out in selected organic solvents to search for the favorable reaction medium. The investigation on reaction parameters showed that optimal laccase activity was obtained in hexane at 30 °C, pH 7.75 for the oxidation of (+)-catechin as well as for (−)-epicatechin, and in toluene at 35 °C, pH 7.25 for the oxidation of catechol. Ea and Q10 values of the biocatalysis in the reaction media of the larger log p solvents like isooctane and hexane were relatively higher than those in the reaction media of lower log p solvents like toluene and dichloromethane. Maximum laccase activity in the organic media was found with 6.5% of buffer as co-solvent. A wider range of 0–28 μg protein/ml in hexane than that of 0–16.7 μg protein/ml in aqueous medium was observed for the linear increasing conversion of (+)-catechin. The kinetic studies revealed that in the presence of isooctane, hexane, toluene and dichloromethane, the Km values were 0.77, 0.97, 0.53 and 2.9 mmol/L for the substrate of (+)-catechin; 0.43, 0.34, 0.14 and 3.4 mmol/L for (−)-epicatechin; 2.9, 1.8, 0.61 and 1.1 mmol/L for catechol, respectively, while the corresponding Vmax values were 2.1 × 10−2, 2.3 × 10−2, 0.65 × 10−2 and 0.71 × 10−2 δA/μg protein min); 1.8 × 10−2, 0.88 × 10−2, 0.19 × 10−2 and 1.0 × 10−2 δA/μg protein min); 0.48 × 10−2, 0.59 × 10−2, 0.67 × 10−2 and 0.54 × 10−2 δA/μg protein min), respectively. FT-IR indicated the formation of probable dimer from (+)-catechin in organic solvent. These results suggest that this laccase has higher catalytic oxidation capacity of phenolic compounds in suitable organic media and favorite oligomers could be obtained.  相似文献   

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