Magnetic Drive Catalytic Application Of Immobilized Hb For The Catalytic Removal Of Phenol

A majority of slaughtering animals blood discarded directly causes mass waste ofslaughter blood in our country. Hemoglobin which contains a kind of hemoprotein haveperoxidase activity. It has dual function such as catalyzing oxidation of phenolcompounds in the existence of hydrogen peroxide, and eliminating toxicity of hydrogenperoxide, phenol, and amine. Based on the research of immobilized Hb technology,utilizing the peroxidase activity of hemoglobin with slaughter blood to handleenvironment wastewater, which removes aromatic compounds in the wastewater, utilizesthe slaughter blood efficiently and meanwhile achieves the purpose of using waste totreat waste.In order to improve the efficiency of the hemoglobin to handle wastewater, thispaper takes the paramagnetism Fe3O4nanoparticles covered by the amino modifiedsilicon as carrier, through glutaraldehyde to make Hb immobilized in single layer on thenanometer carrier. This kind of enzyme preparation can be released, driven and recycledin alternating magnetic field, which makes catalytic reaction of immobilized enzyme outof reaction vessel limitation, avoids the collision among enzymes particles and betweenenzymes particles with reaction vessel, is an ideal process to handle large selection ofwastewater.When the APTS and paramagnetism Fe3O4nanoparticles mass ratio is0.016:5, theimmobilized Hb can reach the best immobilized ratio on the surface of magnetic drivensilicon carrier. When using ABTS as substrates at room temperature without externalmagnetic field, the activity of the immobilized enzyme gets1.7times that of thedissociated enzyme and the immobilized enzyme activity presents quite weaktemperature dependence on enzyme activity changing rule while dissociated enzymepresents typical bell type temperature dependence; under the effect of0-1.2T magneticfield, the catalytic activity of dissociated Hb presents bell type change and under0.4Tmagnetic field its catalytic activity is1.5times that of without magnetic field. Under thefollowing condition of pulse magnetic field: time interval is1.4375s, frequency is0.0625s, intensity between0-0.15T of alternation pulse magnetic field, the catalytic activity of immobilized enzyme presents bell type change, under0.1T magnetic field thecatalytic activity of immobilized enzyme is1.5times that of without magnetic field.Immobilized enzyme has good stability for its oxidation catalyst of enzyme ability tosave more than80%after a long time enzymolysis of pepsin and trypsin enzyme. Thebiological activity of enzyme preparation can keep more than50%after heating30minunder100℃; after reused ten times, enzyme preparation presents good activity stabilityand recovery property (99%); the activity of enzyme preparation preserved in deionizedwater in two months has no obvious change.In research of simulation elimination water sample containing phenol, we found thatunder the following condition to handle the water sample containing phenol throughmagnetic driven immobilized enzyme, phenol content can be achieved from the thirdlevel wastewater of national standard to the first level water:1mM phosphate buffercondition for pH6, phenol concentration for3mg/L, hydrogen peroxide concentrationfor9mM, MAI-Hb concentration for1.0μM, the pulse magnetic field intensity for0.1T,at room temperature (25℃) to react within3hours, phenol clearance rate was98%.The aboved experimental results show that magnetic driven silicon carrier toimmobilize hemoglobin (MAI-Hb) technology have good application prospect in thewastewater containing phenol industry field.