The invention relates to a preparation method of magnetic
chitosan microsphere immobilized
horse radish peroxidases. The preparation method is used in the fields of
enzyme engineering development application and
environmental protection, and
enzyme can be recycled after
enzyme immobilization is carried out. The technical scheme is dissolving
polyethylene glycol 4000in water, adding Fe<2+> salt and Fe<3+> salt according to the
mass ratio of 1:4, dropwise adding stronger
ammonia water until pH of a
system is larger than or equal to 10, adjusting the pH to 7.0 by using
acetic acid after 2h, and conducting
magnetic field separation and washing to obtain magnetofluid; then preparing the
acetic acid solution of
chitosan magnetofluid according to
mass ratio of the
chitosan to the magnetofluid of3:1, ultrasonically blending, adding in atoleine containing emulsifier span 80 for emulsifying, adding
glutaraldehyde for solidification for 3h, and conducting separation, washing and
drying to obtain magnetic
chitosan microspheres; and finally adding the magnetic
chitosan microspheres in horseradish
peroxide enzyme
phosphate buffer, solidifying for 3h at 4DEG C, conducting separation, washing and filtering to prepare the magnetic
chitosan microsphere immobilized
horse radish peroxidases. Carriers of the method are cheap and easy to obtain, enzyme stability is good, and the preparation methodis favorable for rapid and repeated use of the enzyme.