Abstract: In the present study,the effects of immobilized trypsin on small intestinal mucosa were investigated based on the heparin titer.Using heparin titer as the response value,and on the basis of the single factor experimental results,one-factor-at-a-time tests followed by RSM analysis was carried out to achieve the optimal extraction conditions of heparin from porcine intestinal mucosa.The results showed that heparin extraction was significantly affected by pH value,temperature,enzyme dosage,and solid-liquid ratio.The optimal conditions was as follows:enzymatic hydrolysis temperature of 46 ^oC,solid-liquid ratio of 1:3.3 (g/mL),0.24% concentration of immobilized enzyme,pH 11 and 5 hr hydrolysis time,respectively.The heparin titer was reached to 15.28 U under the optimal conditions.

Key words: trypsin, immobilization, heparin, enzyme hydrolysis