Citation:
XIAO Yan, WANG Juan, ZHOU Xiao-Hua. Preparation of Immobilized Chymotrypsin by Chitosan-arginine Resin and Its Properties[J]. Chinese Journal of Applied Chemistry,
;2009, 26(7): 780-785.
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With glutaraldehyde as the crosslinker,chymotrypsin was immobilized onto chitosan-arginine resin which was porous and grafted with a flexible and hydrophilic amino acid.The optimal conditions of immobilization are that the ratio of enzyme loading/wet resin(mg/g) is 20:1,concentration of glutaraldehyde solution is 1.0%,the dose is 10 mL,pH is 5.20,the activity is 850 U/g,and the immobilizing time is 60 min.The enzymatic properties of immobilized chymotrypsin are that the maximum rate is obtained within 30 min,and the catalysis time curve is similar to that of free enzyme;its Km value is 1.83 mmol/L,33.6% higher than that of free enzyme,and 24.0% lower than that of enzyme immobilized by crossing chitosan;its optimal temperature is 70 ℃,10 ℃ higher than that of free enzyme;its optimal pH is 5.92,which deviates 2 units to acid side compared with the free enzyme.Moreover,the immobilized chymotrypsin has a high thermal stability and storage stability,with a half-life of 6 h at 75 ℃ or 49 days at 4 ℃.
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Keywords:
- chitosan,
- L-arginine,
- chymotrypsin,
- cross-linking,
- immobilization
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