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The Effects Of The Support Morphology And The Kinds Of Enzyme Substrate On Mesoporous Carbon CMK-3as Matrix For Horseradish Peroxidase Immobilication

Posted on:2013-10-31Degree:MasterType:Thesis
Country:ChinaCandidate:L WangFull Text:PDF
GTID:2231330395479790Subject:Analytical Chemistry
Abstract/Summary:PDF Full Text Request
The aim of the present work is to explore the effects of the support morphology and thekinds of enzyme substrate on mesoporous materials as matrix for enzyme immobilization insequential injection system. Based on this, the following research experiments wereimplemented by using different morphology mesoporous carbon CMK-3as supports andhorseradish peroxidase(HRP) as model enzyme.Part I Immobilization of horseradish peroxidase on three morphology CMK-3Three different morphologies of CMK-3were served as suppports for the study ofimmobilization of horseradish peroxidase in sequential injection-UV-Vis system. The amountand adsorption mechanisms of horseradish peroxidase on three kinds of morphology ofCMK-3for comparison were studied, and evaluate immobilization HRP activity and stability.Using the nitrogen adsorption and scanning electron microscopy it is found that HRP not onlyadsorb on the surface of CMK-3, but also in the pore, which increase the stability of activityand operation for immobilized horseradish peroxidase.The results show that cosh mesoporous carbon is a fesible immobilization support forhorseradish peroxidase because of the ability of aggregation enzyme or the hydrophobicability and binding capacity is much better than the long stick the rod morphology, so tha ithas the best of immobilization with high activity and good operating stability.Part II immobilization of horseradish peroxidase for specificity selecting of chromogensubstrateThe chromogen substrate of horseradish peroxidase is o-phenylenediamine, guaiacol andcoke phenol, using the sequential injection UV-Vis system for evaluating the substratespecificity.The results prove that the different substrates and products on the CMK-3exist differentadsorbent results, o-phenylenediamine on the support reaching a certain concentration candirectly involved reaction, and coke phenol on the support without adsorption can promote theenzymatic reaction, guaiacol adsorb on the support and product of catalytic reaction havingthe cage-like structure impact on the enzymatic reaction, o-phenylenediamine and cokephenol catalyse H2O2showed a good linear relationship. Furthermore between horseradishperoxidase and coke phenol has more appetency, Km=1.27mmol/L. It is used to identify that that the choice of the substrate is the important factor for reasonable evaluatingimmobilized enzymes...
Keywords/Search Tags:Sequential Injection UV-visible spectrometric system, Immobilized Enzyme, Horseradish peroxide, Mesoporous carbon materials, Chromogen substrat
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