以壳聚糖为载体,戊二醛为交联剂,氨基三乙酸(NTA)为配基,制备固载金属离子的亲和吸附剂,试验了交联剂浓度、配基浓度、偶联温度及时间对配体固载量的影响,最佳条件为戊二醛浓度0.55％、NTA浓度0.4mol·L-1、偶联温度65℃、偶联时间10h.层析分离纯化豆类脲酶,锌柱和镍柱的酶回收率分别为88.5％和80.4％,纯化倍数分别为3.7和6.1,实验结果表明镍柱对脲酶有较高的分离纯化选择性.%Chitosan was used as support,glutaraldehyde as crosslinking agent,nitrilotriacetic acid(NTA)as the chelating ligand.The affinity adsorbent was prepared by immobilized metal ions. The effects on binding amount of ligand of cross-linking agent concentration, genin concentration,binding temperature and time were tested. The optimum condition of the concentration of glutaraldehyde and NTA were 0. 55% and 0.4mol · L-1 respectively,the binding temperature and time were 65% and 10h respectively. Urease from beans was purified by the affinity chromatography. The recovery rate of urease were 88. 5% and 80.4% , the purification fold were 3. 7 and 6. 1 with column chelating Zn( Ⅱ )and Ni( Ⅱ ) respectively. The results showed that affinity chromatography with column chelating Ni( Ⅱ )had a higher selectivity for separation and purification of urease.
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