This paper sets out to summarize the literatures based on immobilized enzyme bio-chromatography and its application in inhibitors screening in the last decade.In order to screen enzyme inhibitors from a mass of compou...This paper sets out to summarize the literatures based on immobilized enzyme bio-chromatography and its application in inhibitors screening in the last decade.In order to screen enzyme inhibitors from a mass of compounds in preliminary screening,multi-pore materials with good biocompatibility are used for the supports of immobilizing enzymes,and then the immobilized enzyme reactor applied as the immobilized enzyme stationary phase in HPLC.Therefore,a technology platform of high throughput screening is gradually established to screen the enzyme inhibitors as new anti-tumor drugs.Here,we briefly summarize the selective methods of supports,immobilization techniques,co-immobilized enzymes system and the screening model.展开更多
Microfluidic,as the systems for using microchannel(micron-or sub-micron scale)to process or manipulate microflow,is being widely applied in enzyme biotechnology and biocatalysis.Microfluidic immobilized enzyme reactor...Microfluidic,as the systems for using microchannel(micron-or sub-micron scale)to process or manipulate microflow,is being widely applied in enzyme biotechnology and biocatalysis.Microfluidic immobilized enzyme reactor(MIER)is a tool with great value for the study of catalytic property and optimal reaction parameter in a flourishing and highly producing manner.In view of its advantages in efficiency,economy,and addressable recognition especially,MIER occupies an important position in the investigation of life science,including molecular biology,bioanalysis and biosensing,biocatalysis etc.Immobilization of enzymes can generally improve their stability,and upon most occasions,the immobilized enzyme is endowed with recyclability.In this review,the enzyme immobilization techniques applied in MIER will be discussed,followed by summarizing the novel developments in the field of MIER for biocatalysis,bioconversion and bioanalysis.The preponderances and deficiencies of the current state-of-the-art preparation ways of MIER are peculiarly discussed.In addition,the prospects of its future study are outlined.展开更多
The synthesis of dipeptide AcPheLeuNH2 catalyzed by immobilized pancreatic lipase was carried out in a two- liquid-phase hollow-fiber membrane reactor, operated in a batch mode. Kinetic properties of free and immobili...The synthesis of dipeptide AcPheLeuNH2 catalyzed by immobilized pancreatic lipase was carried out in a two- liquid-phase hollow-fiber membrane reactor, operated in a batch mode. Kinetic properties of free and immobilized enzyme, partition behavior between aqueous buffer phase and organic solvent phase, and effective diffusion coefficients of substrates and products through the membrane were investigated respectively. Based on the preliminary experimental results, the performance of the enzyme membrane reactor, which is evaluated by the purity and the yield, is discussed.展开更多
A novel strategy for screening the enzyme inhibitors from the natural products by capillary electrophoresis （CE） with a pre column enzyme batch reactor prepared by magnetic microsphere （MB）-gold nanoparticles （Au...A novel strategy for screening the enzyme inhibitors from the natural products by capillary electrophoresis （CE） with a pre column enzyme batch reactor prepared by magnetic microsphere （MB）-gold nanoparticles （AuNPs） is reported. The enzyme batch reactor was prepared by immobilizing the enzymes on the MB conjugated AuNPs （MB-AuNPs）. To demonstrate this strategy, xanthine oxidase （XOD） was employed as a model for the activity of the enzyme, inhibition study, and inhibitor screening. With the developed CE method, the enzyme activity was de- termined by the quantification of the peak area of the product. Enzyme inhibition can be read out directly from the reduced peak area of product in comparison to a reference electropherogram obtained in the absence of any inhibitor A statistical parameter Z＇ factor was recommended for evaluation of the accuracy of a drug screening system. In the present study, it was calculated to be 0.7, implying a good accuracy. The screening of two natural extracts from Cortex Phellodendri and Rhizoma Galangae showed that they were positive for XOD inhibition by the present method. Using this immobilized enzyme technology combined with CE separation not only provides the advantages such as convenience, rapidity and low cost, but also provides a new platform for discovering enzyme-inhibitor drug lead compounds.展开更多
基金supported by the Province Natural Science Foundation of Shandong (Grant number 2009ZRB02230)
文摘This paper sets out to summarize the literatures based on immobilized enzyme bio-chromatography and its application in inhibitors screening in the last decade.In order to screen enzyme inhibitors from a mass of compounds in preliminary screening,multi-pore materials with good biocompatibility are used for the supports of immobilizing enzymes,and then the immobilized enzyme reactor applied as the immobilized enzyme stationary phase in HPLC.Therefore,a technology platform of high throughput screening is gradually established to screen the enzyme inhibitors as new anti-tumor drugs.Here,we briefly summarize the selective methods of supports,immobilization techniques,co-immobilized enzymes system and the screening model.
文摘Microfluidic,as the systems for using microchannel(micron-or sub-micron scale)to process or manipulate microflow,is being widely applied in enzyme biotechnology and biocatalysis.Microfluidic immobilized enzyme reactor(MIER)is a tool with great value for the study of catalytic property and optimal reaction parameter in a flourishing and highly producing manner.In view of its advantages in efficiency,economy,and addressable recognition especially,MIER occupies an important position in the investigation of life science,including molecular biology,bioanalysis and biosensing,biocatalysis etc.Immobilization of enzymes can generally improve their stability,and upon most occasions,the immobilized enzyme is endowed with recyclability.In this review,the enzyme immobilization techniques applied in MIER will be discussed,followed by summarizing the novel developments in the field of MIER for biocatalysis,bioconversion and bioanalysis.The preponderances and deficiencies of the current state-of-the-art preparation ways of MIER are peculiarly discussed.In addition,the prospects of its future study are outlined.
基金Supported by the National Natural Science Foundation of China andExtraction Separation branch of United Chemical Engineeing
文摘The synthesis of dipeptide AcPheLeuNH2 catalyzed by immobilized pancreatic lipase was carried out in a two- liquid-phase hollow-fiber membrane reactor, operated in a batch mode. Kinetic properties of free and immobilized enzyme, partition behavior between aqueous buffer phase and organic solvent phase, and effective diffusion coefficients of substrates and products through the membrane were investigated respectively. Based on the preliminary experimental results, the performance of the enzyme membrane reactor, which is evaluated by the purity and the yield, is discussed.
基金Financial support from the National Natural Science Foundations of China （Nos. 21327007, 21465005）, and the National Basic Research Program of China （No. 211GXNSFDA139006） as well as BAGU1 Scholar Program is gratefully acknowledged.
文摘A novel strategy for screening the enzyme inhibitors from the natural products by capillary electrophoresis （CE） with a pre column enzyme batch reactor prepared by magnetic microsphere （MB）-gold nanoparticles （AuNPs） is reported. The enzyme batch reactor was prepared by immobilizing the enzymes on the MB conjugated AuNPs （MB-AuNPs）. To demonstrate this strategy, xanthine oxidase （XOD） was employed as a model for the activity of the enzyme, inhibition study, and inhibitor screening. With the developed CE method, the enzyme activity was de- termined by the quantification of the peak area of the product. Enzyme inhibition can be read out directly from the reduced peak area of product in comparison to a reference electropherogram obtained in the absence of any inhibitor A statistical parameter Z＇ factor was recommended for evaluation of the accuracy of a drug screening system. In the present study, it was calculated to be 0.7, implying a good accuracy. The screening of two natural extracts from Cortex Phellodendri and Rhizoma Galangae showed that they were positive for XOD inhibition by the present method. Using this immobilized enzyme technology combined with CE separation not only provides the advantages such as convenience, rapidity and low cost, but also provides a new platform for discovering enzyme-inhibitor drug lead compounds.
