Herein,a new nanozyme/natural enzyme hybrid biosensor was established for ultrasensitive l-glutamic acid(L-Glu)detection.The Fe_(3)O_(4) nanoparticles with peroxidase-like activity and stability was used as a nanozyme...Herein,a new nanozyme/natural enzyme hybrid biosensor was established for ultrasensitive l-glutamic acid(L-Glu)detection.The Fe_(3)O_(4) nanoparticles with peroxidase-like activity and stability was used as a nanozyme and carrier for immobilizing l-Glutamate oxidase(GLOD)through Schiffff base reaction to construct a chem-enzyme cascade detector.The resultant Fe_(3)O_(4)-GOLD exhibited a wide linear range(10−500μM)and a low detection limit of 6.03μM for L-Glu detection.Furthermore,the Fe_(3)O_(4)-GOLD exhibited excellent pH stability,thermal stability,reusability and storage stability.After repeated nine cycles,Fe_(3)O_(4)-GOLD still retained 70%of its initial activity.Meanwhile,Fe_(3)O_(4)-GOLD maintained 50%of its initial activity after storage for 20 days,while free GLOD only retained 20%of its initial activity.This strategy of integrating biomimetic Fe 3 O4 and natural enzymes for cascade catalysis makes it possible to design an efficient and stable chemo-enzyme composite catalysts,which are promising for applications in biosensing and biomimetic catalysis.展开更多
The enzyme hybrid nanoflower has gained interests in biosensors due to their simple synthesis and high efficiency.In this study,glutamate oxidase(GLOX)and horseradish peroxidase(HRP)hybrid nanoflowers(GLOX&HRP-HNF...The enzyme hybrid nanoflower has gained interests in biosensors due to their simple synthesis and high efficiency.In this study,glutamate oxidase(GLOX)and horseradish peroxidase(HRP)hybrid nanoflowers(GLOX&HRP-HNFs)were successfully prepared for the detection of glutamic acid(Glu).The effects of the synthesis conditions on the activity of GLOX&HRP-HNFs were investigated.Results revealed that the maximum activity of GLOX&HRP-HNFs was under 4 mM phosphate radical,2.5 mM MnSO4,0.04 mg/mL GLOX,and 0.16 mg/mL HRP.After immobilization,no significant differences were observed in optimum pH and temperature values of the GLOX and HRP.The GLOX&HRP-HNFs exhibited higher storage stability and resistance to organic solvents than free GLOX and HRP.Additionally,the GLOX&HRP-HNFs maintained 69%of its primary activity after 6 cycles.More important,the GLOX&HRP-HNFs exhibited a good linear range from 1 to 100μM(R^(2)=0.9979)and a low limit of detection(LOD)of 0.59μM for glutamate.These results suggest that the GLOX&HRP-HNFs is a promising candidate for applications in biosensing for the detection of glutamate.展开更多
基金supported by the Open Project Program of State Key Laboratory of Food Nutrition and Safety,Tianjin University of Science&Technology(project No.SKLFNS-KF-202203)support from the Science and Technology Program of Tianjin,China(project No.20ZYJDJC00080).
文摘Herein,a new nanozyme/natural enzyme hybrid biosensor was established for ultrasensitive l-glutamic acid(L-Glu)detection.The Fe_(3)O_(4) nanoparticles with peroxidase-like activity and stability was used as a nanozyme and carrier for immobilizing l-Glutamate oxidase(GLOD)through Schiffff base reaction to construct a chem-enzyme cascade detector.The resultant Fe_(3)O_(4)-GOLD exhibited a wide linear range(10−500μM)and a low detection limit of 6.03μM for L-Glu detection.Furthermore,the Fe_(3)O_(4)-GOLD exhibited excellent pH stability,thermal stability,reusability and storage stability.After repeated nine cycles,Fe_(3)O_(4)-GOLD still retained 70%of its initial activity.Meanwhile,Fe_(3)O_(4)-GOLD maintained 50%of its initial activity after storage for 20 days,while free GLOD only retained 20%of its initial activity.This strategy of integrating biomimetic Fe 3 O4 and natural enzymes for cascade catalysis makes it possible to design an efficient and stable chemo-enzyme composite catalysts,which are promising for applications in biosensing and biomimetic catalysis.
基金This work was partially supported by the Open Project Program of State Key Laboratory of Food Nutrition and Safety,Tianjin University of Science&Technology(project No.SKLFNS-KF-202203)Dr.J.D.Cui thanks support from the Science and Technology Program of Tianjin,China(project No.20ZYJDJC00080)the international collaboration project(grant No.2020/37/K/ST8/03805).
文摘The enzyme hybrid nanoflower has gained interests in biosensors due to their simple synthesis and high efficiency.In this study,glutamate oxidase(GLOX)and horseradish peroxidase(HRP)hybrid nanoflowers(GLOX&HRP-HNFs)were successfully prepared for the detection of glutamic acid(Glu).The effects of the synthesis conditions on the activity of GLOX&HRP-HNFs were investigated.Results revealed that the maximum activity of GLOX&HRP-HNFs was under 4 mM phosphate radical,2.5 mM MnSO4,0.04 mg/mL GLOX,and 0.16 mg/mL HRP.After immobilization,no significant differences were observed in optimum pH and temperature values of the GLOX and HRP.The GLOX&HRP-HNFs exhibited higher storage stability and resistance to organic solvents than free GLOX and HRP.Additionally,the GLOX&HRP-HNFs maintained 69%of its primary activity after 6 cycles.More important,the GLOX&HRP-HNFs exhibited a good linear range from 1 to 100μM(R^(2)=0.9979)and a low limit of detection(LOD)of 0.59μM for glutamate.These results suggest that the GLOX&HRP-HNFs is a promising candidate for applications in biosensing for the detection of glutamate.