A new method of the preparation of carrier of enzyme immobilization magnetic polyacrylonitrile microsphere was reported in this paper. Dispersed the nanometer magnetic fluid into the N,N dimethylformamide solution of polyacrylonitrile, the suparamagnetic polyacrylonitrile microsphere was obtained by using spray method. The influence factor on the particle properties were investigated in detail. Under the optimum conditions: C PAN =1g/L, T =180℃, n =30 000r/min, V =2mL/s, C magnetic fluid =2g/L, suparamagnetic polyacrylonitrile microspheres with narrow disperse(1.0μm~1.5μm) were prepared.