A new nanocomposite material for construction of glucose biosensor was prepared. The biosensor was formed by entrapping glucose oxidase(Gox) into chitosan/nanoporous ZrO2/multiwalled carbon nanotubes nanocomposite fil...A new nanocomposite material for construction of glucose biosensor was prepared. The biosensor was formed by entrapping glucose oxidase(Gox) into chitosan/nanoporous ZrO2/multiwalled carbon nanotubes nanocomposite film. In this biosensing thin film, the multiwalled carbon nanotubes can effectively catalyze hydrogen peroxide and nanoporous ZrO2 can enhance the stability of the immobilized enzyme. The resulting biosensor provides a very effective matrix for the immobilization of glucose oxidase and exhibits a wide linear response range from 8 μmol/L to 3 mmol/L with a correlation coefficient of 0.994 for the detection of glucose. And the response time and detection limit of the biosensor are determined to be 6 s and 3.5 μmol/L, respectively. Another attractive characteristic is that the biosensor is inexpensive, stable and reliable.展开更多
Raw corn starch granules were hydrolysized by glucoamylase in a chemostat. The hydro- lysis of three different-sized granules shows that smaller granules undergo more hydrolyzation than larger ones. After 78 h, 9700 o...Raw corn starch granules were hydrolysized by glucoamylase in a chemostat. The hydro- lysis of three different-sized granules shows that smaller granules undergo more hydrolyzation than larger ones. After 78 h, 9700 of the granules was hydrolysized with diameter between 0.15 mm and 0.3 mm at 50 ℃. When corn starch concentration increased from 100 g/L to 250 g/L, the amount of reducing sugar produced was proportional to the initial substrate concentration and no substrate inhibition phenomenon appeared. In order to study the product inhibition exactly, the product from hydrolysis reaction itself was added into the hydrolysis system at the beginning of starch hydrolysis. Product inhibition with different quantities of product added were studied in the initial several hours, during which period enzyme inactivation could be neglected and product inhibition could be studied separately. The experiments indicate that product inhibition happens when the additional quantity exceeds 9.56 g/L.展开更多
A novel polyaniline-graphite composite film glucose oxidase (PGCF GOD) electrode was developed. The PGCF was synthesized by cyclic voitammetry method in 0.5 mol/L H2SO4 solution containing 1 g/L graphite powder and ...A novel polyaniline-graphite composite film glucose oxidase (PGCF GOD) electrode was developed. The PGCF was synthesized by cyclic voitammetry method in 0.5 mol/L H2SO4 solution containing 1 g/L graphite powder and 0.2 mol/L aniline. The PGCF GOD electrode was prepared by doping GOD into the composite film. The morphology of the PGCF and the response property of the PGCF GOD electrode were investigated by scanning electron microscopy and electrochemical measurement, respectively. The results show that the PGCF has a porous and netty structure and the PGCF GOD electrode has excellent response property such as high sensitivity and short response time. Influences of pH value, temperature, glucose concentration and potential on the response current of the electrode were also discussed. The sensor has a maximum steady-state current density of 357.17μA/cm2 and an apparent Michaelis-Menten constant of 16.57 mmol/L. The maximum current response of the enzyme electrode occurs under the condition ofpH 5.5, 0.8 V and 65℃.展开更多
ZnO nanorods, with the c-axis orientation used for transparent conductors, solar cells, sensors especially the functionalized ZnO nanorods with some kinds of enzymes have been used for biosensor. In this work, we desc...ZnO nanorods, with the c-axis orientation used for transparent conductors, solar cells, sensors especially the functionalized ZnO nanorods with some kinds of enzymes have been used for biosensor. In this work, we describe the process immobilization of galactose oxidase on ZnO nanorods surface with glutaraldehyde as a cross-linker molecule to make the working electrode in electrochemical biosensor. ZnO nanorods were grown on FTO (Fluorine-doped tin oxide) substrate by solution method at low temperature. The crystalline phase and orientation of ZnO nanorods were identified using X-ray diffraction. The efficiency of the immobilization was calculated by Braford method showed that about 36% enzyme content was immobilized on ZnO nanorods surface. The working electrode based on the immobilized ZnO nanorods was tested in galactose solution by CV (cyclic voltammetry) method indicated the value of current intensity is about 0.14 μA. These results clearly demonstrate the potential of galactose sensor based on ZnO nanorod.展开更多
Invertase hydrolyses sucrose, produces inverted sugar syrup, which is used, mainly, as a food composition in industries. To carry out the hydrolysis properly, the invertase should be stable in the soluble form through...Invertase hydrolyses sucrose, produces inverted sugar syrup, which is used, mainly, as a food composition in industries. To carry out the hydrolysis properly, the invertase should be stable in the soluble form through a considerable reaction period and recovered afterwards. The chosen reactor was a CSTR-type (continuous stirred tank reactor-type) coupled with an UFM (ultrafiltration-membrane), the so-called MR (membrane reactor). The varied parameters were: sucrose concentration (10-300 mM), temperature (5-65 ℃), reaction volume (14 mL and 65 mL), stirring (100-500 rpm), volumetric feeding rate (2.2-12 mL/h) and UFM MWCO (molecular weight cut off) (10, 20, 30, 50 and 100 kDa). The invertase kinetic constants (KM = 23.5 mM; Vmax = 2,758 μmolgluJmin-mgprot; Ea = 9.1 kcal/mol) and the temperature deactivation energy (Ead= 20 kcal/mol) were calculated. Moreover, the invertase was unstable as the MR capacity diminished and the agitation increased up to 500 rpm most likely due to the damaging effect of shearing forces (present inside the MR) on the invertase molecule. Finally, both the MWCO and the chemical nature of the UFM affected the invertase stability along the hydrolysis. The enzyme stability increased as the UFM cut-off decreased, the highest value being observed with the 10 kDa-UFM.展开更多
A novel electrochemiluminescence (ECL) sensing approach was developed for glucose detection based on crosslinking Ru(bpy)3Cl2-doped silica nanoparticles (RuSiNPs) with glucose dehydrogenase on a glassy carbon electrod...A novel electrochemiluminescence (ECL) sensing approach was developed for glucose detection based on crosslinking Ru(bpy)3Cl2-doped silica nanoparticles (RuSiNPs) with glucose dehydrogenase on a glassy carbon electrode (GCE). Glutaral- dehyde and aminopropyltrimethoxysilane were used as linking agents, and chitosan was used to immobilize the composites onto the GCE surface. The ECL sensor presented good characteristics in terms of stability and reproducibility. Under opti- mized conditions, the linear response of ECL intensity to glucose concentration was valid in the range of 0.2 to 20 mmol/L (R2 = 0.9962). The application results indicated that the proposed approach is with great potential in the determination of glucose.展开更多
The platinum nanoparticles were adsorbed on graphene oxide sheets and played an important role in catalytic reduction of graphene oxide with hydrazine, leading to the formation of graphene-Pt nanoparticles. Because of...The platinum nanoparticles were adsorbed on graphene oxide sheets and played an important role in catalytic reduction of graphene oxide with hydrazine, leading to the formation of graphene-Pt nanoparticles. Because of their good electronic properties, biocompatibility and high surface area, graphene-Pt based composites achieved the direct electron transfer of redox enzyme and maintained their bioactivity well. The graphene-Pt nanocomposites were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HR-TEM) and selected area electron diffraction (SAED). The amperometric biosensor fabricated by depositing glucose oxidase over Nafion-solubilized graphene-Pt electrode retained its biocatalytic activity and has offered fast and sensitive glucose quantification.展开更多
Chemiluminescence detection was developed as an alternative to amperometric detection for glucose analysis in a portable, microfluidicsbased continuous glucose monitoring system. Amperometric detection allows easy det...Chemiluminescence detection was developed as an alternative to amperometric detection for glucose analysis in a portable, microfluidicsbased continuous glucose monitoring system. Amperometric detection allows easy determination of hydrogen peroxide, a product of the glucose oxidasecatalyzed reaction of glucose with oxygen, by oxidation at a microelectrode. However, (micro)electrodes in direct contact with physiological sample are subject to electrode fouling, which leads to signal drift, decreased reproducibility and shortened detector lifetimes. Moreover, there are a few species present in the body (e.g. ascorbic acid, uric acid) which can undergo oxidation at the same applied potential as hydrogen peroxide. These species can thus inter- fere with the glucose measurement, reducing detection specificity. The rationale for exploring chemiluminescence as opposed to amperometric detection is thus to attempt to improve the lifetime and reproducibility of glucose analysis for monitoring purposes, while reducing interference caused by other chemicals in the body. The study reported here represents a first step in this direction, namely the realization of a microfluidic device with integrated silicon photodiode for chemiluminescence detection of glucose. This microflow device uses a chaotic mixing approach to perform enzymatic conversion of glucose, followed by reaction of the hydrogen peroxide produced with luminol to produce light at 425 nm. The chemil reaction is catalyzed by horseradish peroxidase in the presence of iodophenol. The performance of the fabricated chip was characterized to establish optimal reaction conditions with respect to sample and reagent flow rates, pH, and concentrations. A linear calibra- tion curve was obtained for current response as a function of glucose concentration in the clinically relevant range between 2 and 10 mM, with a sensitivity of 39 pA/mM (R = 0.9963, one device, n = 3) and a limit of detection of 230 ktM (S/N - 3).展开更多
The paper reports a novel amperometric biosensor for catechol based on immobilization of a highly sensitive horseradish peroxidase by affinity interactions on metal chelate-functionalized agarose/carbon nanotubes comp...The paper reports a novel amperometric biosensor for catechol based on immobilization of a highly sensitive horseradish peroxidase by affinity interactions on metal chelate-functionalized agarose/carbon nanotubes composites. Metal chelate affinity takes advantage of the affinity of Ni2+ ions to bind strongly and reversibly to histidine or cysteine tails found on the surface of the horseradish peroxidase. Thus, enzymes with such residues in their molecules can be easily attached to functionalized aga- rose/carbon nanotubes composites support containing a nickel chelate. Linear sweep voltammograms and amperometry are used to study the proposed electrochemical biosensor. Catechol is determined by direct reduction of biocatalytically liberated quinone species at -0.05 V (vs. SCE). The effect ofpH, applied electrode potential and the concentration of H2O2 on the sensitivity of the biosensor has been investigated. The performance of the proposed biosensor is tested using four different phenolic compounds, showing very high sensitivity, in particular, the linearity of cateehol is observed from 2.0 × 10-8 to 1.05×10-5 M with a detection limit of 5.0×10-9 M.展开更多
基金Project (20060532006) supported by Specialized Research Fund for the Doctoral Program of Higher Education
文摘A new nanocomposite material for construction of glucose biosensor was prepared. The biosensor was formed by entrapping glucose oxidase(Gox) into chitosan/nanoporous ZrO2/multiwalled carbon nanotubes nanocomposite film. In this biosensing thin film, the multiwalled carbon nanotubes can effectively catalyze hydrogen peroxide and nanoporous ZrO2 can enhance the stability of the immobilized enzyme. The resulting biosensor provides a very effective matrix for the immobilization of glucose oxidase and exhibits a wide linear response range from 8 μmol/L to 3 mmol/L with a correlation coefficient of 0.994 for the detection of glucose. And the response time and detection limit of the biosensor are determined to be 6 s and 3.5 μmol/L, respectively. Another attractive characteristic is that the biosensor is inexpensive, stable and reliable.
文摘Raw corn starch granules were hydrolysized by glucoamylase in a chemostat. The hydro- lysis of three different-sized granules shows that smaller granules undergo more hydrolyzation than larger ones. After 78 h, 9700 of the granules was hydrolysized with diameter between 0.15 mm and 0.3 mm at 50 ℃. When corn starch concentration increased from 100 g/L to 250 g/L, the amount of reducing sugar produced was proportional to the initial substrate concentration and no substrate inhibition phenomenon appeared. In order to study the product inhibition exactly, the product from hydrolysis reaction itself was added into the hydrolysis system at the beginning of starch hydrolysis. Product inhibition with different quantities of product added were studied in the initial several hours, during which period enzyme inactivation could be neglected and product inhibition could be studied separately. The experiments indicate that product inhibition happens when the additional quantity exceeds 9.56 g/L.
基金Projects(50473022, 20673036) supported by the National Natural Science Foundation of China project(2005) supported by the State Key Laboratory of Chemo/Biosensing and Chemometrics of China+1 种基金 project(2006FJ4100) supported by the Science Technology Project of Hunan Province project(2006) supported by the Postdoctor Foundation of Hunan University
文摘A novel polyaniline-graphite composite film glucose oxidase (PGCF GOD) electrode was developed. The PGCF was synthesized by cyclic voitammetry method in 0.5 mol/L H2SO4 solution containing 1 g/L graphite powder and 0.2 mol/L aniline. The PGCF GOD electrode was prepared by doping GOD into the composite film. The morphology of the PGCF and the response property of the PGCF GOD electrode were investigated by scanning electron microscopy and electrochemical measurement, respectively. The results show that the PGCF has a porous and netty structure and the PGCF GOD electrode has excellent response property such as high sensitivity and short response time. Influences of pH value, temperature, glucose concentration and potential on the response current of the electrode were also discussed. The sensor has a maximum steady-state current density of 357.17μA/cm2 and an apparent Michaelis-Menten constant of 16.57 mmol/L. The maximum current response of the enzyme electrode occurs under the condition ofpH 5.5, 0.8 V and 65℃.
文摘ZnO nanorods, with the c-axis orientation used for transparent conductors, solar cells, sensors especially the functionalized ZnO nanorods with some kinds of enzymes have been used for biosensor. In this work, we describe the process immobilization of galactose oxidase on ZnO nanorods surface with glutaraldehyde as a cross-linker molecule to make the working electrode in electrochemical biosensor. ZnO nanorods were grown on FTO (Fluorine-doped tin oxide) substrate by solution method at low temperature. The crystalline phase and orientation of ZnO nanorods were identified using X-ray diffraction. The efficiency of the immobilization was calculated by Braford method showed that about 36% enzyme content was immobilized on ZnO nanorods surface. The working electrode based on the immobilized ZnO nanorods was tested in galactose solution by CV (cyclic voltammetry) method indicated the value of current intensity is about 0.14 μA. These results clearly demonstrate the potential of galactose sensor based on ZnO nanorod.
文摘Invertase hydrolyses sucrose, produces inverted sugar syrup, which is used, mainly, as a food composition in industries. To carry out the hydrolysis properly, the invertase should be stable in the soluble form through a considerable reaction period and recovered afterwards. The chosen reactor was a CSTR-type (continuous stirred tank reactor-type) coupled with an UFM (ultrafiltration-membrane), the so-called MR (membrane reactor). The varied parameters were: sucrose concentration (10-300 mM), temperature (5-65 ℃), reaction volume (14 mL and 65 mL), stirring (100-500 rpm), volumetric feeding rate (2.2-12 mL/h) and UFM MWCO (molecular weight cut off) (10, 20, 30, 50 and 100 kDa). The invertase kinetic constants (KM = 23.5 mM; Vmax = 2,758 μmolgluJmin-mgprot; Ea = 9.1 kcal/mol) and the temperature deactivation energy (Ead= 20 kcal/mol) were calculated. Moreover, the invertase was unstable as the MR capacity diminished and the agitation increased up to 500 rpm most likely due to the damaging effect of shearing forces (present inside the MR) on the invertase molecule. Finally, both the MWCO and the chemical nature of the UFM affected the invertase stability along the hydrolysis. The enzyme stability increased as the UFM cut-off decreased, the highest value being observed with the 10 kDa-UFM.
基金financially supported by the Science and Technology Project of Hebei Province
文摘A novel electrochemiluminescence (ECL) sensing approach was developed for glucose detection based on crosslinking Ru(bpy)3Cl2-doped silica nanoparticles (RuSiNPs) with glucose dehydrogenase on a glassy carbon electrode (GCE). Glutaral- dehyde and aminopropyltrimethoxysilane were used as linking agents, and chitosan was used to immobilize the composites onto the GCE surface. The ECL sensor presented good characteristics in terms of stability and reproducibility. Under opti- mized conditions, the linear response of ECL intensity to glucose concentration was valid in the range of 0.2 to 20 mmol/L (R2 = 0.9962). The application results indicated that the proposed approach is with great potential in the determination of glucose.
基金supported by the National Natural Science Foundation of China (Grant No. 21003151)the Fundamental Research Funds for the Central Universities (Grant No. 0400219212)
文摘The platinum nanoparticles were adsorbed on graphene oxide sheets and played an important role in catalytic reduction of graphene oxide with hydrazine, leading to the formation of graphene-Pt nanoparticles. Because of their good electronic properties, biocompatibility and high surface area, graphene-Pt based composites achieved the direct electron transfer of redox enzyme and maintained their bioactivity well. The graphene-Pt nanocomposites were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HR-TEM) and selected area electron diffraction (SAED). The amperometric biosensor fabricated by depositing glucose oxidase over Nafion-solubilized graphene-Pt electrode retained its biocatalytic activity and has offered fast and sensitive glucose quantification.
文摘Chemiluminescence detection was developed as an alternative to amperometric detection for glucose analysis in a portable, microfluidicsbased continuous glucose monitoring system. Amperometric detection allows easy determination of hydrogen peroxide, a product of the glucose oxidasecatalyzed reaction of glucose with oxygen, by oxidation at a microelectrode. However, (micro)electrodes in direct contact with physiological sample are subject to electrode fouling, which leads to signal drift, decreased reproducibility and shortened detector lifetimes. Moreover, there are a few species present in the body (e.g. ascorbic acid, uric acid) which can undergo oxidation at the same applied potential as hydrogen peroxide. These species can thus inter- fere with the glucose measurement, reducing detection specificity. The rationale for exploring chemiluminescence as opposed to amperometric detection is thus to attempt to improve the lifetime and reproducibility of glucose analysis for monitoring purposes, while reducing interference caused by other chemicals in the body. The study reported here represents a first step in this direction, namely the realization of a microfluidic device with integrated silicon photodiode for chemiluminescence detection of glucose. This microflow device uses a chaotic mixing approach to perform enzymatic conversion of glucose, followed by reaction of the hydrogen peroxide produced with luminol to produce light at 425 nm. The chemil reaction is catalyzed by horseradish peroxidase in the presence of iodophenol. The performance of the fabricated chip was characterized to establish optimal reaction conditions with respect to sample and reagent flow rates, pH, and concentrations. A linear calibra- tion curve was obtained for current response as a function of glucose concentration in the clinically relevant range between 2 and 10 mM, with a sensitivity of 39 pA/mM (R = 0.9963, one device, n = 3) and a limit of detection of 230 ktM (S/N - 3).
基金supported by the National Outstanding Youth Foundations of China (50725825)National Basic Research Program of China (2007CB310501 & 2011CB935704)+2 种基金National Natural Science Foundation of China (50908113)the Natural Science Foundation of Jiangxi Province (2008GZH0008)the Youth Foundation of Jiangxi Provincial Department of Education (GJJ09483)
文摘The paper reports a novel amperometric biosensor for catechol based on immobilization of a highly sensitive horseradish peroxidase by affinity interactions on metal chelate-functionalized agarose/carbon nanotubes composites. Metal chelate affinity takes advantage of the affinity of Ni2+ ions to bind strongly and reversibly to histidine or cysteine tails found on the surface of the horseradish peroxidase. Thus, enzymes with such residues in their molecules can be easily attached to functionalized aga- rose/carbon nanotubes composites support containing a nickel chelate. Linear sweep voltammograms and amperometry are used to study the proposed electrochemical biosensor. Catechol is determined by direct reduction of biocatalytically liberated quinone species at -0.05 V (vs. SCE). The effect ofpH, applied electrode potential and the concentration of H2O2 on the sensitivity of the biosensor has been investigated. The performance of the proposed biosensor is tested using four different phenolic compounds, showing very high sensitivity, in particular, the linearity of cateehol is observed from 2.0 × 10-8 to 1.05×10-5 M with a detection limit of 5.0×10-9 M.